RNA interference in dermal and fibrotic indications

ABSTRACT

The present invention relates to RNAi constructs with improved tissue and cellular uptake characteristics and methods of use of these compounds in dermal and fibrotic applications.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/636,755, entitled “RNA INTERFERENCE IN DERMAL AND FIBROTICINDICATIONS”, filed Apr. 4, 2013, which is a National Stage Applicationof PCT/US2011/029867, entitled “RNA INTERFERENCE IN DERMAL AND FIBROTICINDICATIONS,” filed on Mar. 24, 2011, which was published under PCTArticle 21(2) in English and which claims the benefit under 35 U.S.C. §119(e) of U.S. 61/317,252, entitled “RNA INTERFERENCE IN SKININDICATIONS,” filed on Mar. 24, 2010, and U.S. Provisional ApplicationSer. No. U.S. 61/317,633, entitled “RNA INTERFERENCE IN SKININDICATIONS,” filed on Mar. 25, 2010, each of which is hereinincorporated by reference in its entirety.

FIELD OF INVENTION

The invention pertains to the field of RNA interference (RNAi). Theinvention more specifically relates to nucleic acid molecules withimproved in vivo delivery properties and their use for dermal andfibrotic indications.

BACKGROUND OF INVENTION

Complementary oligonucleotide sequences are promising therapeutic agentsand useful research tools in elucidating gene functions. However, priorart oligonucleotide molecules suffer from several problems that mayimpede their clinical development, and frequently make it difficult toachieve intended efficient inhibition of gene expression (includingprotein synthesis) using such compositions in vivo.

A major problem has been the delivery of these compounds to cells andtissues. Conventional double-stranded RNAi compounds, 19-29 bases long,form a highly negatively-charged rigid helix of approximately 1.5 by10-15 nm in size. This rod type molecule cannot get through thecell-membrane and as a result has very limited efficacy both in vitroand in vivo. As a result, all conventional RNAi compounds require somekind of a delivery vehicle to promote their tissue distribution andcellular uptake. This is considered to be a major limitation of the RNAitechnology.

There have been previous attempts to apply chemical modifications tooligonucleotides to improve their cellular uptake properties. One suchmodification was the attachment of a cholesterol molecule to theoligonucleotide. A first report on this approach was by Letsinger etal., in 1989. Subsequently, ISIS Pharmaceuticals, Inc. (Carlsbad,Calif.) reported on more advanced techniques in attaching thecholesterol molecule to the oligonucleotide (Manoharan, 1992).

With the discovery of siRNAs in the late nineties, similar types ofmodifications were attempted on these molecules to enhance theirdelivery profiles. Cholesterol molecules conjugated to slightly modified(Soutschek, 2004) and heavily modified (Wolfrum, 2007) siRNAs appearedin the literature. Yamada et al., 2008 also reported on the use ofadvanced linker chemistries which further improved cholesterol mediateduptake of siRNAs. In spite of all this effort, the uptake of these typesof compounds appears to be inhibited in the presence of biologicalfluids resulting in highly limited efficacy in gene silencing in vivo,limiting the applicability of these compounds in a clinical setting.

SUMMARY OF INVENTION

Described herein is the efficient in vivo delivery of sd-rxRNA moleculesto the skin and the use of such molecules for gene silencing. This classof RNAi molecules has superior efficacy both in vitro and in vivo thanpreviously described RNAi molecules. Molecules associated with theinvention have widespread potential as therapeutics for disorders orconditions associated with compromised skin and fibrosis.

Aspects of the invention relate to double-stranded ribonucleic acids(dsRNAs) including a sense strand and an antisense strand wherein theantisense strand is complementary to at least 12 contiguous nucleotidesof a sequence selected from the sequences within Tables 2, 5, 6, 9, 11,12, 13, 14, 15, 16, 17 and 23, and wherein the dsRNA is an sd-rxRNA.

Further aspects of the invention relate to double-stranded ribonucleicacids (dsRNAs) comprising a sense strand and an antisense strand whereinthe sense strand and/or the antisense strand comprises at least 12contiguous nucleotides of a sequence selected from the sequences withinTables 1-27, and wherein the dsRNA is an sd-rxRNA.

Further aspects of the invention relate to double-stranded ribonucleicacids (dsRNAs) comprising a sense strand and an antisense strand whereinthe antisense strand is complementary to at least 12 contiguousnucleotides of a sequence selected from the sequences within Tables 2,5, 6, 9, 11, 12, 13, 14, 15, 16, 17 and 23, and wherein the dsRNA is anrxRNAori.

Further aspects of the invention relate to double-stranded ribonucleicacids (dsRNAs) comprising a sense strand and an antisense strand whereinthe sense strand and/or the antisense strand comprises at least 12contiguous nucleotides of a sequence selected from the sequences withinTables 1-27, and wherein the dsRNA is an rxRNAori.

In some embodiments, the dsRNA is directed against CTGF. In someembodiments, the antisense strand of the dsRNA is complementary to atleast 12 contiguous nucleotides of a sequence selected from thesequences within Tables 11, 12 and 15. In some embodiments, the sensestrand and/or the antisense strand comprises at least 12 contiguousnucleotides of a sequence selected from the sequences within Tables 10,11, 12, 15, 20 and 24.

In some embodiments, the sense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: SEQ IDNOs: 2463, 3429, 2443, 3445, 2459, 3493, 2465 and 3469. In someembodiments, the antisense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: 2464,3430, 4203, 3446, 2460, 3494, 2466 and 3470.

In certain embodiments, the sense strand comprises SEQ ID NO:2463 andthe antisense strand comprises SEQ ID NO:2464. In certain embodiments,the sense strand comprises SEQ ID NO:3429 and the antisense strandcomprises SEQ ID NO:3430.

In certain embodiments, the sense strand comprises SEQ ID NO:2443 andthe antisense strand comprises SEQ ID NO:4203. In certain embodiments,the sense strand comprises SEQ ID NO:3445 and the antisense strandcomprises SEQ ID NO:3446.

In certain embodiments, the sense strand comprises SEQ ID NO:2459 andthe antisense strand comprises SEQ ID NO:2460. In certain embodiments,the sense strand comprises SEQ ID NO:3493 and the antisense strandcomprises SEQ ID NO:3494.

In certain embodiments, the sense strand comprises SEQ ID NO:2465 andthe antisense strand comprises SEQ ID NO:2466. In certain embodiments,the sense strand comprises SEQ ID NO:3469 and the antisense strandcomprises SEQ ID NO:3470.

In some embodiments, the sense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: SEQ IDNOs: 1835, 1847, 1848 and 1849. In certain embodiments, the sense strandcomprises a sequence selected from the group consisting of: SEQ ID NOs:1835, 1847, 1848 and 1849.

In some embodiments, the dsRNA is hydrophobically modified. In certainembodiments, the dsRNA is linked to a hydrophobic conjugate.

Aspects of the invention relate to compositions comprising the dsRNAdescribed herein. In some embodiments, the composition comprises dsRNAdirected against genes encoding for more than one protein.

In some embodiments, the composition is formulated for delivery to theskin. In certain embodiments, the composition is in a neutralformulation. In some embodiments, the composition is formulated fortopical delivery or for intradermal injection.

Aspects of the invention relate to methods comprising delivering any ofthe dsRNA described herein or a composition comprising any of the dsRNAdescribed herein to the skin of a subject in need thereof.

Aspects of the invention relate to methods comprising administering to asubject in need thereof a therapeutically effective amount of a doublestranded ribonucleic acid (dsRNA) comprising a sense strand and anantisense strand wherein the antisense strand is complementary to atleast 12 contiguous nucleotides of a sequence selected from thesequences within Tables 2, 5, 6, 9, 11, 12, 13, 14, 15, 16, 17 and 23,and wherein the dsRNA is an sd-rxRNA.

Further aspects of the invention relate to methods comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a double stranded ribonucleic acid (dsRNA) comprising a sensestrand and an antisense strand wherein the sense strand and/or theantisense strand comprises at least 12 contiguous nucleotides of asequence selected from the sequences within Tables 1-27, and wherein thedsRNA is an sd-rxRNA.

Further aspects of the invention relate to methods comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a double stranded ribonucleic acid (dsRNA) comprising a sensestrand and an antisense strand wherein the antisense strand iscomplementary to at least 12 contiguous nucleotides of a sequenceselected from the sequences within Tables 2, 5, 6, 9, 11, 12, 13, 14,15, 16, 17 and 23, and wherein the dsRNA is an rxRNAori.

Further aspects of the invention relate to methods comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a double stranded ribonucleic acid (dsRNA) comprising a sensestrand and an antisense strand wherein the sense strand and/or theantisense strand comprises at least 12 contiguous nucleotides of asequence selected from the sequences within Tables 1-27, and wherein thedsRNA is an rxRNAori.

In some embodiments, the method is a method for treating compromisedskin. In some embodiments, the method is a method for treating orpreventing a fibrotic disorder.

In some embodiments, the dsRNA is administered via intradermalinjection. In some embodiments, the dsRNA is administered locally to theskin. In some embodiments, two or more nucleic acid molecules areadministered simultaneously or sequentially.

In some embodiments, one or more of the dsRNAs is hydrophobicallymodified. In certain embodiments, one or more of the dsRNAs is linked toa hydrophobic conjugate.

In some embodiments, the dsRNA is directed against CTGF. In certainembodiments, the antisense strand of the dsRNA is complementary to atleast 12 contiguous nucleotides of a sequence selected from thesequences within Tables 11, 12 and 15. In some embodiments, the sensestrand and/or the antisense strand comprises at least 12 contiguousnucleotides of a sequence selected from the sequences within Tables 10,11, 12, 15, 20 and 24.

In some embodiments, the sense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: SEQ IDNOs: 2463, 3429, 2443, 3445, 2459, 3493, 2465 and 3469. In certainembodiments, the antisense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: 2464,3430, 4203, 3446, 2460, 3494, 2466 and 3470.

In certain embodiments, the sense strand comprises SEQ ID NO:2463 andthe antisense strand comprises SEQ ID NO:2464. In certain embodiments,the sense strand comprises SEQ ID NO:3429 and the antisense strandcomprises SEQ ID NO:3430.

In certain embodiments, the sense strand comprises SEQ ID NO:2443 andthe antisense strand comprises SEQ ID NO:4203. In certain embodiments,the sense strand comprises SEQ ID NO:3445 and the antisense strandcomprises SEQ ID NO:3446.

In certain embodiments, the sense strand comprises SEQ ID NO:2459 andthe antisense strand comprises SEQ ID NO:2460. In certain embodiments,the sense strand comprises SEQ ID NO:3493 and the antisense strandcomprises SEQ ID NO:3494.

In certain embodiments, the sense strand comprises SEQ ID NO:2465 andthe antisense strand comprises SEQ ID NO:2466. In certain embodiments,the sense strand comprises SEQ ID NO:3469 and the antisense strandcomprises SEQ ID NO:3470.

In some embodiments, the sense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: SEQ IDNOs: 1835, 1847, 1848 and 1849. In some embodiments, the sense strandcomprises a sequence selected from the group consisting of: SEQ ID NOs:1835, 1847, 1848 and 1849.

Aspects of the invention relate to treating or preventing a fibroticdisorder. In some embodiments, the fibrotic disorder is selected fromthe group consisting of pulmonary fibrosis, liver cirrhosis, sclerodermaand glomerulonephritis, lung fibrosis, liver fibrosis, skin fibrosis,muscle fibrosis, radiation fibrosis, kidney fibrosis, proliferativevitreoretinopathy, restenosis and uterine fibrosis, and trabeculectomyfailure due to scarring.

In some embodiments, the dsRNA are administered via intradermalinjection, while in other embodiments, the one or more dsRNA areadministered subcutaneously or epicutaneously.

The one or more dsRNA can be administered prior to, during and/or aftera medical procedure. In some embodiments, administration occurs within 8days prior to or within 8 days after the medical procedure. In someembodiments, the medical procedure is surgery. In certain embodiments,the surgery is elective. In some embodiments, the surgery comprisesepithelial grafting or skin grafting. In some embodiments, the one ormore double stranded nucleic acid molecules are administered to a graftdonor site and/or a graft recipient site.

Aspects of the invention relate to methods for administering one or moredsRNA prior to, during and/or after an injury. In some embodiments, thesubject has a wound such as a chronic wound. In certain embodiments, thewound is a result of elective surgery. The wound can be external orinternal. In some embodiments, the dsRNA is administered after burninjury.

Methods described herein include methods for promoting wound healing andmethods for preventing scarring.

In some embodiments, one or more of the dsRNA administered to a subjectis directed against a gene selected from the group consisting of TGFB1,TGFB2, hTGFB1, hTGFB2, PTGS2, SPP1, hSPP1, CTGF or hCTGF. In someembodiments, the one or more dsRNA are administered on the skin of thesubject. In certain embodiments, the one or more dsRNA molecules are inthe form of a cream or ointment. In some embodiments, two or more orthree or more nucleic acids are administered. Two or more nucleic acidmolecules can be administered simultaneously or sequentially.

Aspects of the invention related to nucleic acids that are optimized. Insome embodiments, one or more double stranded nucleic acid molecules arehydrophobically modified. In certain embodiments, the one or more doublestranded nucleic acid molecules are linked to a hydrophobic conjugate ormultiple hydrophobic conjugates. In some embodiments, the one or moredouble stranded nucleic acid molecule are linked to a lipophilic group.In certain embodiments, the lipophilic group is linked to the passengerstrand of the one or more double stranded nucleic acid molecules. Insome embodiment, the one or more double stranded nucleic acid moleculesare linked to cholesterol, a long chain alkyl cholesterol analog,vitamin A or vitamin E. In some embodiments, the one or more doublestranded nucleic acid molecules is attached to chloroformate.

Aspects of the invention related to nucleic acids that are optimizedthrough modifications. In some embodiments, the one or more doublestranded nucleic acid molecules includes at least one 2′ O methyl or 2′fluoro modification and/or at least one 5 methyl C or U modification. Insome embodiments, the one or more double stranded nucleic acid moleculeshas a guide strand of 16-28 nucleotides in length. In certainembodiments, at least 40% of the nucleotides of the one or more doublestranded nucleic acid molecules are modified. Double stranded nucleicacid molecules described herein can also be attached to linkers. In someembodiments, the linker is protonatable.

Aspects of the invention relate to double stranded nucleic acidmolecules that contain at least two single stranded regions. In someembodiments, the single stranded regions contain phosphorothioatemodifications. In certain embodiments, the single stranded regions arelocated at the 3′ end of the guide strand and the 5′ end of thepassenger strand.

Aspects of the invention relate to methods for delivering a nucleic acidto a subject, involving administering to a subject within 8 days priorto a medical procedure a therapeutically effective amount for treatingcompromised skin of one or more sd-rxRNAs.

Each of the limitations of the invention can encompass variousembodiments of the invention. It is, therefore, anticipated that each ofthe limitations of the invention involving any one element orcombinations of elements can be included in each aspect of theinvention. This invention is not limited in its application to thedetails of construction and the arrangement of components set forth inthe following description or illustrated in the drawings. The inventionis capable of other embodiments and of being practiced or of beingcarried out in various ways.

BRIEF DESCRIPTION OF DRAWINGS

The accompanying drawings are not intended to be drawn to scale. In thedrawings, each identical or nearly identical component that isillustrated in various figures is represented by a like numeral. Forpurposes of clarity, not every component may be labeled in everydrawing. In the drawings:

FIG. 1 demonstrates the expression profiles for non-limiting examples oftarget genes including MAP4K4, SPP1, CTGF, PTGS2 and TGFB1. As expected,target gene expression is elevated early and returns to normal by day10.

FIG. 2 presents schematics depicting an experimental approach tovisualizing tissue after intradermal injection.

FIG. 3 demonstrates silencing of MAP4K4 following intradermal injectionof sd-rxRNA targeting MAP4K4.

FIG. 4 demonstrates silencing of MAP4K4, PPIB and CTGF followingintradermal injection of sd-rxRNA molecules targeting each gene.

FIG. 5 demonstrates silencing of MAP4K4 following intradermal injectionof sd-rxRNA targeting MAP4K4. Normalized expression of MAP4K4 relativeto controls is demonstrated.

FIG. 6 demonstrates silencing of PPIB following intradermal injection ofsd-rxRNA targeting PPIB. Normalized expression of PPIB relative tocontrols is demonstrated.

FIG. 7 demonstrates the duration of PPIB silencing following intradermalinjection of sd-rxRNA targeting PPIB.

FIG. 8 demonstrates the duration of MAP4K4 silencing followingintradermal injection of sd-rxRNA targeting MAP4K4.

FIG. 9 demonstrates equivalent silencing achieved using two differentdosing regimens.

FIG. 10 demonstrates examples of sd-rxRNA molecules targeting CTGF thatare efficacious for gene silencing.

FIG. 11 demonstrates examples of sd-rxRNA molecules targeting CTGF thatare efficacious for gene silencing.

FIG. 12 demonstrates a dose response for sd-rxRNA molecules targetingCTGF.

FIG. 13 demonstrates a sample of an original sd-rxRNA screen.

FIG. 14 presents data on a hit from the original sd-rxRNA screen.

FIG. 15 demonstrates gene expression of PTGS2 following administrationof sd-rxRNA targeting PTGS2.

FIG. 16 demonstrates gene expression of hTGFB1 following administrationof sd-rxRNA targeting hTGFB1.

FIG. 17 demonstrates gene expression of hTGFB1 following administrationof sd-rxRNA targeting hTGFB1.

FIG. 18 demonstrates results of TGFB1 sd-rxRNA screening.

FIG. 19 demonstrates gene expression of TGFB2 following administrationof sd-rxRNA targeting TGFB2.

FIG. 20 demonstrates gene expression of TGFB2 following administrationof sd-rxRNA targeting TGFB2.

FIG. 21 demonstrates gene expression of TGFB2 following administrationof sd-rxRNA targeting TGFB2.

FIG. 22 demonstrates gene expression of TGFB2 following administrationof sd-rxRNA targeting TGFB2.

FIG. 23 demonstrates gene expression of TGFB2 following administrationof sd-rxRNA targeting TGFB2.

FIG. 24 demonstrates results of TGFB2 sd-rxRNA screening.

FIG. 25 demonstrates identification of potent hSPP1 sd-rxRNAs.

FIG. 26 demonstrates identification of potent hSPP1 sd-rxRNAs.

FIG. 27 demonstrates identification of potent hSPP1 sd-rxRNAs.

FIG. 28 demonstrates SPP1 sd-rxRNA compound selection.

FIG. 29 demonstrates that variation of linker chemistry does notinfluence silencing activity of sd-rxRNAs in vitro. Two different linkerchemistries were evaluated, a hydroxyproline linker and ribo linker, onmultiple sd-rxRNAs (targeting Map4k4 or PPIB) in passive uptake assaysto determine linkers which favor self delivery. HeLa cells weretransfected in the absence of a delivery vehicle (passive transfection)with sd-rxRNAs at 1 uM, 0.1 uM or 0.01 uM for 48 hrs. Use of eitherlinker results in an efficacious delivery of sd-rxRNA.

FIG. 30 depicts CTGF as a central factor in the pathway to fibrosis.

FIG. 31 depicts the phases of wound healing.

FIG. 32 depicts the chemical optimization of sd-rxRNA leads.

FIG. 33 demonstrates that chemically optimized CTGF L1 sd-rxRNAs areactive.

FIG. 34 demonstrates in vitro efficacy of chemically optimized CTGF L1sd-rxRNAs.

FIG. 35 demonstrates in vitro stability of chemically optimized CTGF L1sd-rxRNAs.

FIG. 36 demonstrates that chemically optimized CTGF L2 sd-rxRNAs areactive.

FIG. 37 demonstrates in vitro efficacy of chemically optimized CTGF L2sd-rxRNAs.

FIG. 38 demonstrates in vitro stability of chemically optimized CTGF L2sd-rxRNAs.

FIG. 39 provides a summary of compounds that are active in vivo.

FIG. 40 demonstrates that treatment with CTGF L1B target sequenceresulted in mRNA silencing.

FIG. 41 demonstrates that treatment with CTGF L2 target sequenceresulted in mRNA silencing.

FIG. 42 demonstrates CTGF silencing after two intradermal injections ofRXi-109.

FIG. 43 demonstrates the duration of CTGF silencing in skin afterintradermal injection of the sd-rxRNA in SD rats. Eight millimeter skinbiopsies were harvested, and mRNA levels were quantified by QPCR andnormalized to a housekeeping gene. Shown is percent (%) silencing vs.Non Targeting Control (NTC); PBS at each time point is one experimentalgroup; * p≤0.04; ** p≤0.002.

FIG. 44 demonstrates that chemically optimized CTGF L3 sd-rxRNAs areactive.

FIG. 45 demonstrates absolute luminescence of CTGF L4 sd-rxRNAs.

FIG. 46 demonstrates that chemically optimized CTGF L4 sd-rxRNAs areactive.

FIG. 47 demonstrates changes in mRNA expression levels of CTGF, α-SMactin, collagen 1A2, and collagen 3A1 after intradermal injection ofCTFG sd-rxRNA in SD rats. mRNA levels were quantified by qPCR.

FIG. 48 demonstrates that there is no apparent delay in wound healingwith treatment of CTGF-targeting sd-rxRNA. Some changes was observedwith treatment of a combination of CTGF- and COX2-targeting sd-rxRNAs.

FIG. 49 demonstrates that administration of sd-rxRNAs decreases woundwidth over the course of at least 9 days. The graph shows microscopicmeasurements of wound width in rats on days 3, 6, and 9 post-wounding.Each group represents 5 rats. Two non-serial sections from each woundwere measured and the average width of the two was calculated per wound.*p<0.05 vs. PBS an NTC.

FIG. 50 demonstrates that administration of sd-rxRNAs decreases woundarea over the course of at least 9 days. The graph shows microscopicmeasurements of wound width in rats on days 3, 6, and 9 post-wounding.Each group represents 5 rats. Two non-serial sections from each woundwere measured and the average width of the two was calculated per wound.*p<0.05 vs. PBS an NTC.

FIG. 51 demonstrates that administration of sd-rxRNAs increase thepercentage of wound re-epithelialization over the course of at least 9days. The graph shows microscopic measurements of wound width in rats ondays 3, 6, and 9 post-wounding. Each group represents 5 rats. Twonon-serial sections from each wound were measured and the average widthof the two was calculated per wound. *p<0.05 vs. PBS an NTC.

FIG. 52 demonstrates that administration of sd-rxRNAs increases theaverage granulation tissue maturity scores over the course of at least 9days. The graph shows microscopic measurements of wound width in rats ondays 3, 6, and 9 post-wounding (5=mature, 1=immature). Each grouprepresents 5 rats.

FIG. 53 demonstrates CD68 labeling in day 9 wounds (0=no labeling,3=substantial labeling). Each group represents 5 rats.

FIG. 54 demonstrates that CTGF leads have different toxicity levels invitro.

FIG. 55 shows percentage (%) of cell viability after RXI 109 doseescalation (oligos formulated in PBS).

FIG. 56 is a schematic of Phases 1 and 2 clinical trial design.

FIG. 57 is a schematic of Phases 1 and 2 clinical trial design.

FIG. 58 demonstrates a percent (%) decrease in PPIB expression in theliver relative to PBS control. Lipoid formulated rxRNAs (10 mg/kg) weredelivery systemically to Balb/c mice (n=5) by single tail veininjections. Liver tissue was harvested at 24 hours after injection andexpression was analyzed by qPCR (normalized to β-actin). Map4K4 rxRNAorialso showed significant silencing (˜83%, p<0.001) although Map4K4sd-rxRNA did not significantly reduce target gene expression (˜17%,p=0.019). TD.035.2278, Published lipidoid delivery reagent, 98N12-5(1),from Akinc, 2009.

FIG. 59 demonstrates that chemically optimized PTGS2 L1 sd-rxRNAs areactive.

FIG. 60 demonstrates that chemically optimized PTGS2 L2 sd-rxRNAs areactive.

FIG. 61 demonstrates that chemically optimized hTGFB1 L1 sd-rxRNAs areactive.

FIG. 62 demonstrates that chemically optimized hTGFB1 L1 sd-rxRNAs areactive.

FIG. 63 demonstrates that chemically optimized hTGFB2 L1 sd-rxRNAs areactive.

FIG. 64 demonstrates that chemically optimized hTGFB2 sd-rxRNAs areactive.

DETAILED DESCRIPTION

Aspects of the invention relate to methods and compositions involved ingene silencing. The invention is based at least in part on thesurprising discovery that administration of sd-rxRNA molecules to theskin, such as through intradermal injection or subcutaneousadministration, results in efficient silencing of gene expression in theskin. Highly potent sd-rxRNA molecules that target genes including SPP1,CTGF, PTGS2, TGFB1 and TGFB2 were also identified herein throughcell-based screening. sd-rxRNAs represent a new class of therapeuticRNAi molecules with significant potential in treatment of compromisedskin.

sd-rxRNA Molecules

Aspects of the invention relate to sd-rxRNA molecules. As used herein,an “sd-rxRNA” or an “sd-rxRNA molecule” refers to a self-delivering RNAmolecule such as those described in, and incorporated by reference from,PCT Publication No. WO2010/033247 (Application No. PCT/US2009/005247),filed on Sep. 22, 2009, and entitled “REDUCED SIZE SELF-DELIVERING RNAICOMPOUNDS,” and PCT application PCT/US2009/005246, filed on Sep. 22,2009, and entitled “RNA INTERFERENCE IN SKIN INDICATIONS.” Briefly, ansd-rxRNA, (also referred to as an sd-rxRNA^(nano)) is an isolatedasymmetric double stranded nucleic acid molecule comprising a guidestrand, with a minimal length of 16 nucleotides, and a passenger strandof 8-18 nucleotides in length, wherein the double stranded nucleic acidmolecule has a double stranded region and a single stranded region, thesingle stranded region having 4-12 nucleotides in length and having atleast three nucleotide backbone modifications. In preferred embodiments,the double stranded nucleic acid molecule has one end that is blunt orincludes a one or two nucleotide overhang. sd-rxRNA molecules can beoptimized through chemical modification, and in some instances throughattachment of hydrophobic conjugates.

In some embodiments, an sd-rxRNA comprises an isolated double strandednucleic acid molecule comprising a guide strand and a passenger strand,wherein the region of the molecule that is double stranded is from 8-15nucleotides long, wherein the guide strand contains a single strandedregion that is 4-12 nucleotides long, wherein the single stranded regionof the guide strand contains 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12phosphorothioate modifications, and wherein at least 40% of thenucleotides of the double stranded nucleic acid are modified.

The polynucleotides of the invention are referred to herein as isolateddouble stranded or duplex nucleic acids, oligonucleotides orpolynucleotides, nano molecules, nano RNA, sd-rxRNA^(nano), sd-rxRNA orRNA molecules of the invention.

sd-rxRNAs are much more effectively taken up by cells compared toconventional siRNAs. These molecules are highly efficient in silencingof target gene expression and offer significant advantages overpreviously described RNAi molecules including high activity in thepresence of serum, efficient self delivery, compatibility with a widevariety of linkers, and reduced presence or complete absence of chemicalmodifications that are associated with toxicity.

In contrast to single-stranded polynucleotides, duplex polynucleotideshave traditionally been difficult to deliver to a cell as they haverigid structures and a large number of negative charges which makesmembrane transfer difficult. sd-rxRNAs however, although partiallydouble-stranded, are recognized in vivo as single-stranded and, as such,are capable of efficiently being delivered across cell membranes. As aresult the polynucleotides of the invention are capable in manyinstances of self delivery. Thus, the polynucleotides of the inventionmay be formulated in a manner similar to conventional RNAi agents orthey may be delivered to the cell or subject alone (or with non-deliverytype carriers) and allowed to self deliver. In one embodiment of thepresent invention, self delivering asymmetric double-stranded RNAmolecules are provided in which one portion of the molecule resembles aconventional RNA duplex and a second portion of the molecule is singlestranded.

The oligonucleotides of the invention in some aspects have a combinationof asymmetric structures including a double stranded region and a singlestranded region of 5 nucleotides or longer, specific chemicalmodification patterns and are conjugated to lipophilic or hydrophobicmolecules. This class of RNAi like compounds have superior efficacy invitro and in vivo. It is believed that the reduction in the size of therigid duplex region in combination with phosphorothioate modificationsapplied to a single stranded region contribute to the observed superiorefficacy.

The invention is based at least in part on the surprising discovery thatsd-rxRNA molecules are delivered efficiently in vivo to the skin througha variety of methods including intradermal injection and subcutaneousadministration. Furthermore, sd-rxRNA molecules are efficient inmediating gene silencing in the region of the skin where they aretargeted.

Aspects of the invention relate to the use of cell-based screening toidentify potent sd-rxRNA molecules. Described herein is theidentification of potent sd-rxRNA molecules that target a subset ofgenes including SPP1, CTFG, PTGS2, TGFB1 and TGFB2. In some embodiments,a target gene is selected and an algorithm is applied to identifyoptimal target sequences within that gene (Example 2). For example, manysequences can be selected for one gene. In some instances, the sequencesthat are identified are generated as RNAi compounds for a first round oftesting. For example, the RNAi compounds based on the optimal predictedsequences can initially be generated as rxRNAori (“ori”) sequences forthe first round of screening. After identifying potent RNAi compounds,these can be generated as sd-rxRNA molecules.

dsRNA formulated according to the invention also includes rxRNAori.rxRNAori refers to a class of RNA molecules described in andincorporated by reference from PCT Publication No. WO2009/102427(Application No. PCT/US2009/000852), filed on Feb. 11, 2009, andentitled, “MODIFIED RNAI POLYNUCLEOTIDES AND USES THEREOF.”

In some embodiments, an rxRNAori molecule comprises a double-strandedRNA (dsRNA) construct of 12-35 nucleotides in length, for inhibitingexpression of a target gene, comprising: a sense strand having a 5′-endand a 3′-end, wherein the sense strand is highly modified with2′-modified ribose sugars, and wherein 3-6 nucleotides in the centralportion of the sense strand are not modified with 2′-modified ribosesugars and, an antisense strand having a 5′-end and a 3′-end, whichhybridizes to the sense strand and to mRNA of the target gene, whereinthe dsRNA inhibits expression of the target gene in a sequence-dependentmanner.

rxRNAori can contain any of the modifications described herein. In someembodiments, at least 30% of the nucleotides in the rxRNAori aremodified. For example, at least 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%,38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%,52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%,66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% of the nucleotides in the rxRNAori aremodified. In some embodiments, 100% of the nucleotides in the sd-rxRNAare modified. In some embodiments, only the passenger strand of therxRNAori contains modifications.

In some embodiments, the RNAi compounds of the invention comprise anasymmetric compound comprising a duplex region (required for efficientRISC entry of 8-15 bases long) and single stranded region of 4-12nucleotides long; with a 13 or 14 nucleotide duplex. A 6 or 7 nucleotidesingle stranded region is preferred in some embodiments. The singlestranded region of the new RNAi compounds also comprises 2-12phosphorothioate internucleotide linkages (referred to asphosphorothioate modifications). 6-8 phosphorothioate internucleotidelinkages are preferred in some embodiments. Additionally, the RNAicompounds of the invention also include a unique chemical modificationpattern, which provides stability and is compatible with RISC entry. Thecombination of these elements has resulted in unexpected propertieswhich are highly useful for delivery of RNAi reagents in vitro and invivo.

The chemical modification pattern, which provides stability and iscompatible with RISC entry includes modifications to the sense, orpassenger, strand as well as the antisense, or guide, strand. Forinstance the passenger strand can be modified with any chemical entitieswhich confirm stability and do not interfere with activity. Suchmodifications include 2′ ribo modifications (O-methyl, 2′ F, 2 deoxy andothers) and backbone modification like phosphorothioate modifications. Apreferred chemical modification pattern in the passenger strand includesOmethyl modification of C and U nucleotides within the passenger strandor alternatively the passenger strand may be completely Omethylmodified.

The guide strand, for example, may also be modified by any chemicalmodification which confirms stability without interfering with RISCentry. A preferred chemical modification pattern in the guide strandincludes the majority of C and U nucleotides being 2′ F modified and the5′ end being phosphorylated. Another preferred chemical modificationpattern in the guide strand includes 2′ Omethyl modification of position1 and C/U in positions 11-18 and 5′ end chemical phosphorylation. Yetanother preferred chemical modification pattern in the guide strandincludes 2′Omethyl modification of position 1 and C/U in positions 11-18and 5′ end chemical phosphorylation and 2′F modification of C/U inpositions 2-10. In some embodiments the passenger strand and/or theguide strand contains at least one 5-methyl C or U modifications.

In some embodiments, at least 30% of the nucleotides in the sd-rxRNA aremodified. For example, at least 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%,38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%,52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%,66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% of the nucleotides in the sd-rxRNA aremodified. In some embodiments, 100% of the nucleotides in the sd-rxRNAare modified.

The above-described chemical modification patterns of theoligonucleotides of the invention are well tolerated and actuallyimproved efficacy of asymmetric RNAi compounds.

It was also demonstrated experimentally herein that the combination ofmodifications to RNAi when used together in a polynucleotide results inthe achievement of optimal efficacy in passive uptake of the RNAi.Elimination of any of the described components (Guide strandstabilization, phosphorothioate stretch, sense strand stabilization andhydrophobic conjugate) or increase in size in some instances results insub-optimal efficacy and in some instances complete lost of efficacy.The combination of elements results in development of a compound, whichis fully active following passive delivery to cells such as HeLa cells.

The data in the Examples presented below demonstrates high efficacy ofthe oligonucleotides of the invention both in vitro in variety of celltypes and in vivo upon local and systemic administration.

The sd-rxRNA can be further improved in some instances by improving thehydrophobicity of compounds using of novel types of chemistries. Forexample one chemistry is related to use of hydrophobic basemodifications. Any base in any position might be modified, as long asmodification results in an increase of the partition coefficient of thebase. The preferred locations for modification chemistries are positions4 and 5 of the pyrimidines. The major advantage of these positions is(a) ease of synthesis and (b) lack of interference with base-pairing andA form helix formation, which are essential for RISC complex loading andtarget recognition. A version of sd-rxRNA compounds where multiple deoxyUridines are present without interfering with overall compound efficacywas used. In addition major improvement in tissue distribution andcellular uptake might be obtained by optimizing the structure of thehydrophobic conjugate. In some of the preferred embodiment the structureof sterol is modified to alter (increase/decrease) C17 attached chain.This type of modification results in significant increase in cellularuptake and improvement of tissue uptake prosperities in vivo.

Aspects of the invention relate to double-stranded ribonucleic acidmolecules (dsRNA) such as sd-rxRNA and rxRNAori. dsRNA associated withthe invention can comprise a sense strand and an antisense strandwherein the antisense strand is complementary to at least 12 contiguousnucleotides of a sequence selected from the sequences within Tables 2,5, 6, 9, 11, 12, 13, 14, 15, 16, 17 and 23. For example, the antisensestrand can be complementary to at least 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, or 24 contiguous nucleotides, or can be complementary to25 nucleotides of a sequence selected from the sequences within Tables2, 5, 6, 9, 11, 12, 13, 14, 15, 16, 17 and 23.

dsRNA associated with the invention can comprise a sense strand and anantisense strand wherein the sense strand and/or the antisense strandcomprises at least 12 contiguous nucleotides of a sequence selected fromthe sequences within Tables 1-27. For example, the sense strand and/orthe antisense strand can comprise at least 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, or 24 contiguous nucleotides, or can comprise 25nucleotides of a sequence selected from the sequences within Tables1-27.

Aspects of the invention relate to dsRNA directed against CTGF. Forexample, the antisense strand of a dsRNA directed against CTGF can becomplementary to at least 12 contiguous nucleotides of a sequenceselected from the sequences within Tables 11, 12 and 15. The sensestrand and/or the antisense strand of a dsRNA directed against CTGF cancomprises at least 12 contiguous nucleotides of a sequence selected fromthe sequences within Tables 10, 11, 12, 15, 20 and 24.

In some embodiments, the sense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting of: SEQ IDNOs: 2463, 3429, 2443, 3445, 2459, 3493, 2465 and 3469. In certainembodiments, the sense strand comprises or consists of a sequenceselected from the group consisting of: SEQ ID NOs: 2463, 3429, 2443,3445, 2459, 3493, 2465 and 3469.

In some embodiments, the antisense strand comprises at least 12contiguous nucleotides of a sequence selected from the group consistingof: 2464, 3430, 4203, 3446, 2460, 3494, 2466 and 3470. In certainembodiments, the antisense strand comprises or consists of a sequenceselected from the group consisting of: 2464, 3430, 4203, 3446, 2460,3494, 2466 and 3470.

In a preferred embodiment, the sense strand comprises SEQ ID NO:2463(GCACCUUUCUAGA) and the antisense strand comprises SEQ ID NO:2464(UCUAGAAAGGUGCAAACAU). The sequences of SEQ ID NO:2463 and SEQ IDNO:2464 can be modified in a variety of ways according to modificationsdescribed herein. A preferred modification pattern for SEQ ID NO:2463 isdepicted by SEQ ID NO:3429 (G.mC. A.mC.mC.mU.mU.mU.mC.mU.A*mG*mA.TEG-Chl). A preferred modification pattern for SEQ ID NO:2464 isdepicted by SEQ ID NO:3430 (P.mU.fC.fU. A. G.mA. A.mA. G. G.fU. G.mC* A*A* A*mC* A* U). An sd-rxRNA consisting of SEQ ID NO:3429 and SEQ IDNO:3430 is also referred to as RXi-109.

In another preferred embodiment, the sense strand comprises SEQ IDNO:2443 (UUGCACCUUUCUAA) and the antisense strand comprises SEQ IDNO:4203 (UUAGAAAGGUGCAAACAAGG). The sequences of SEQ ID NO:2443 and SEQID NO:4203 can be modified in a variety of ways according tomodifications described herein. A preferred modification pattern for SEQID NO:2443 is depicted by SEQ ID NO:3445 (mU.mU. G.mC.A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl). A preferred modification patternfor SEQ ID NO:4203 is depicted by SEQ ID NO:3446 (P.mU.fU. A. G. A.mA.A. G. G.fU. G.fC.mA.mA*mA*fC*mA*mA*mG* G.).

In another preferred embodiment, the sense strand comprises SEQ IDNO:2459 (GUGACCAAAAGUA) and the antisense strand comprises SEQ IDNO:2460 (UACUUUUGGUCACACUCUC). The sequences of SEQ ID NO:2459 and SEQID NO:2460 can be modified in a variety of ways according tomodifications described herein. A preferred modification pattern for SEQID NO:2459 is depicted by SEQ ID NO:3493 (G.mU. G. A.mC.mC. A. A. A. A.G*mU*mA.TEG-Chl). A preferred modification pattern for SEQ ID NO:2460 isdepicted by SEQ ID NO:3494 (P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC*A*mC*mU*mC*mU* C.).

In another preferred embodiment, the sense strand comprises SEQ IDNO:2465 (CCUUUCUAGUUGA) and the antisense strand comprises SEQ IDNO:2466 (UCAACUAGAAAGGUGCAAA). The sequences of SEQ ID NO:2465 and SEQID NO:2466 can be modified in a variety of ways according tomodifications described herein. A preferred modification pattern for SEQID NO:2465 is depicted by SEQ ID NO:3469 (mC.mC.mU.mU.mU.mC.mU. A.G.mU.mU*mG*mA.TEG-Chl). A preferred modification pattern for SEQ IDNO:2466 is depicted by SEQ ID NO:3470 (P.mU.fC. A. A.fC.fU. A. G. A.mA.A. G. G*fU*mG*fC*mA*mA* A.).

A preferred embodiment of an rxRNAori directed against CTGF can compriseat least 12 contiguous nucleotides of a sequence selected from the groupconsisting of: SEQ ID NOs:1835, 1847, 1848 and 1849. In someembodiments, the sense strand of the rxRNAori comprises or consists ofSEQ ID NOs:1835, 1847, 1848 or 1849.

Aspects of the invention relate to compositions comprising dsRNA such assd-rxRNA and rxRNAori. In some embodiments compositions comprise two ormore dsRNA that are directed against different genes.

This invention is not limited in its application to the details ofconstruction and the arrangement of components set forth in thefollowing description or illustrated in the drawings. The invention iscapable of other embodiments and of being practiced or of being carriedout in various ways. Also, the phraseology and terminology used hereinis for the purpose of description and should not be regarded aslimiting. The use of “including,” “comprising,” or “having,”“containing,” “involving,” and variations thereof herein, is meant toencompass the items listed thereafter and equivalents thereof as well asadditional items.

Thus, aspects of the invention relate to isolated double strandednucleic acid molecules comprising a guide (antisense) strand and apassenger (sense) strand. As used herein, the term “double-stranded”refers to one or more nucleic acid molecules in which at least a portionof the nucleomonomers are complementary and hydrogen bond to form adouble-stranded region. In some embodiments, the length of the guidestrand ranges from 16-29 nucleotides long. In certain embodiments, theguide strand is 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or29 nucleotides long. The guide strand has complementarity to a targetgene. Complementarity between the guide strand and the target gene mayexist over any portion of the guide strand. Complementarity as usedherein may be perfect complementarity or less than perfectcomplementarity as long as the guide strand is sufficientlycomplementary to the target that it mediates RNAi. In some embodimentscomplementarity refers to less than 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%,or 1% mismatch between the guide strand and the target. Perfectcomplementarity refers to 100% complementarity. Thus the invention hasthe advantage of being able to tolerate sequence variations that mightbe expected due to genetic mutation, strain polymorphism, orevolutionary divergence. For example, siRNA sequences with insertions,deletions, and single point mutations relative to the target sequencehave also been found to be effective for inhibition. Moreover, not allpositions of a siRNA contribute equally to target recognition.Mismatches in the center of the siRNA are most critical and essentiallyabolish target RNA cleavage. Mismatches upstream of the center orupstream of the cleavage site referencing the antisense strand aretolerated but significantly reduce target RNA cleavage. Mismatchesdownstream of the center or cleavage site referencing the antisensestrand, preferably located near the 3′ end of the antisense strand, e.g.1, 2, 3, 4, 5 or 6 nucleotides from the 3′ end of the antisense strand,are tolerated and reduce target RNA cleavage only slightly.

While not wishing to be bound by any particular theory, in someembodiments, the guide strand is at least 16 nucleotides in length andanchors the Argonaute protein in RISC. In some embodiments, when theguide strand loads into RISC it has a defined seed region and targetmRNA cleavage takes place across from position 10-11 of the guidestrand. In some embodiments, the 5′ end of the guide strand is or isable to be phosphorylated. The nucleic acid molecules described hereinmay be referred to as minimum trigger RNA.

In some embodiments, the length of the passenger strand ranges from 8-15nucleotides long. In certain embodiments, the passenger strand is 8, 9,10, 11, 12, 13, 14 or 15 nucleotides long. The passenger strand hascomplementarity to the guide strand. Complementarity between thepassenger strand and the guide strand can exist over any portion of thepassenger or guide strand. In some embodiments, there is 100%complementarity between the guide and passenger strands within thedouble stranded region of the molecule.

Aspects of the invention relate to double stranded nucleic acidmolecules with minimal double stranded regions. In some embodiments theregion of the molecule that is double stranded ranges from 8-15nucleotides long. In certain embodiments, the region of the moleculethat is double stranded is 8, 9, 10, 11, 12, 13, 14 or 15 nucleotideslong. In certain embodiments the double stranded region is 13 or 14nucleotides long. There can be 100% complementarity between the guideand passenger strands, or there may be one or more mismatches betweenthe guide and passenger strands. In some embodiments, on one end of thedouble stranded molecule, the molecule is either blunt-ended or has aone-nucleotide overhang. The single stranded region of the molecule isin some embodiments between 4-12 nucleotides long. For example thesingle stranded region can be 4, 5, 6, 7, 8, 9, 10, 11 or 12 nucleotideslong. However, in certain embodiments, the single stranded region canalso be less than 4 or greater than 12 nucleotides long. In certainembodiments, the single stranded region is 6 nucleotides long.

RNAi constructs associated with the invention can have a thermodynamicstability (ΔG) of less than −13 kkal/mol. In some embodiments, thethermodynamic stability (ΔG) is less than −20 kkal/mol. In someembodiments there is a loss of efficacy when (ΔG) goes below −21kkal/mol. In some embodiments a (ΔG) value higher than −13 kkal/mol iscompatible with aspects of the invention. Without wishing to be bound byany theory, in some embodiments a molecule with a relatively higher (ΔG)value may become active at a relatively higher concentration, while amolecule with a relatively lower (ΔG) value may become active at arelatively lower concentration. In some embodiments, the (ΔG) value maybe higher than −9 kkcal/mol. The gene silencing effects mediated by theRNAi constructs associated with the invention, containing minimal doublestranded regions, are unexpected because molecules of almost identicaldesign but lower thermodynamic stability have been demonstrated to beinactive (Rana et al. 2004).

Without wishing to be bound by any theory, results described hereinsuggest that a stretch of 8-10 bp of dsRNA or dsDNA will be structurallyrecognized by protein components of RISC or co-factors of RISC.Additionally, there is a free energy requirement for the triggeringcompound that it may be either sensed by the protein components and/orstable enough to interact with such components so that it may be loadedinto the Argonaute protein. If optimal thermodynamics are present andthere is a double stranded portion that is preferably at least 8nucleotides then the duplex will be recognized and loaded into the RNAimachinery.

In some embodiments, thermodynamic stability is increased through theuse of LNA bases. In some embodiments, additional chemical modificationsare introduced. Several non-limiting examples of chemical modificationsinclude: 5′ Phosphate, 2′-O-methyl, 2′-O-ethyl, 2′-fluoro,ribothymidine, C-5 propynyl-dC (pdC) and C-5 propynyl-dU (pdU); C-5propynyl-C (pC) and C-5 propynyl-U (pU); 5-methyl C, 5-methyl U,5-methyl dC, 5-methyl dU methoxy, (2,6-diaminopurine),5′-Dimethoxytrityl-N4-ethyl-2′-deoxyCytidine and MGB (minor groovebinder). It should be appreciated that more than one chemicalmodification can be combined within the same molecule.

Molecules associated with the invention are optimized for increasedpotency and/or reduced toxicity. For example, nucleotide length of theguide and/or passenger strand, and/or the number of phosphorothioatemodifications in the guide and/or passenger strand, can in some aspectsinfluence potency of the RNA molecule, while replacing 2′-fluoro (2′F)modifications with 2′-O-methyl (2′OMe) modifications can in some aspectsinfluence toxicity of the molecule. Specifically, reduction in 2′Fcontent of a molecule is predicted to reduce toxicity of the molecule.The Examples section presents molecules in which 2′F modifications havebeen eliminated, offering an advantage over previously described RNAicompounds due to a predicted reduction in toxicity. Furthermore, thenumber of phosphorothioate modifications in an RNA molecule caninfluence the uptake of the molecule into a cell, for example theefficiency of passive uptake of the molecule into a cell. Preferredembodiments of molecules described herein have no 2′F modification andyet are characterized by equal efficacy in cellular uptake and tissuepenetration. Such molecules represent a significant improvement overprior art, such as molecules described by Accell and Wolfrum, which areheavily modified with extensive use of 2′F.

In some embodiments, a guide strand is approximately 18-19 nucleotidesin length and has approximately 2-14 phosphate modifications. Forexample, a guide strand can contain 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14 or more than 14 nucleotides that are phosphate-modified. Theguide strand may contain one or more modifications that confer increasedstability without interfering with RISC entry. The phosphate modifiednucleotides, such as phosphorothioate modified nucleotides, can be atthe 3′ end, 5′ end or spread throughout the guide strand. In someembodiments, the 3′ terminal 10 nucleotides of the guide strand contains1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 phosphorothioate modified nucleotides.The guide strand can also contain 2′F and/or 2′OMe modifications, whichcan be located throughout the molecule. In some embodiments, thenucleotide in position one of the guide strand (the nucleotide in themost 5′ position of the guide strand) is 2′OMe modified and/orphosphorylated. C and U nucleotides within the guide strand can be 2′Fmodified. For example, C and U nucleotides in positions 2-10 of a 19 ntguide strand (or corresponding positions in a guide strand of adifferent length) can be 2′F modified. C and U nucleotides within theguide strand can also be 2′OMe modified. For example, C and Unucleotides in positions 11-18 of a 19 nt guide strand (or correspondingpositions in a guide strand of a different length) can be 2′OMemodified. In some embodiments, the nucleotide at the most 3′ end of theguide strand is unmodified. In certain embodiments, the majority of Csand Us within the guide strand are 2′F modified and the 5′ end of theguide strand is phosphorylated. In other embodiments, position 1 and theCs or Us in positions 11-18 are 2′OMe modified and the 5′ end of theguide strand is phosphorylated. In other embodiments, position 1 and theCs or Us in positions 11-18 are 2′OMe modified, the 5′ end of the guidestrand is phosphorylated, and the Cs or Us in position 2-10 are 2′Fmodified.

In some aspects, an optimal passenger strand is approximately 11-14nucleotides in length. The passenger strand may contain modificationsthat confer increased stability. One or more nucleotides in thepassenger strand can be 2′OMe modified. In some embodiments, one or moreof the C and/or U nucleotides in the passenger strand is 2′OMe modified,or all of the C and U nucleotides in the passenger strand are 2′OMemodified. In certain embodiments, all of the nucleotides in thepassenger strand are 2′OMe modified. One or more of the nucleotides onthe passenger strand can also be phosphate-modified such asphosphorothioate modified. The passenger strand can also contain 2′ribo, 2′F and 2 deoxy modifications or any combination of the above. Asdemonstrated in the Examples, chemical modification patterns on both theguide and passenger strand are well tolerated and a combination ofchemical modifications is shown herein to lead to increased efficacy andself-delivery of RNA molecules.

Aspects of the invention relate to RNAi constructs that have extendedsingle-stranded regions relative to double stranded regions, as comparedto molecules that have been used previously for RNAi. The singlestranded region of the molecules may be modified to promote cellularuptake or gene silencing. In some embodiments, phosphorothioatemodification of the single stranded region influences cellular uptakeand/or gene silencing. The region of the guide strand that isphosphorothioate modified can include nucleotides within both the singlestranded and double stranded regions of the molecule. In someembodiments, the single stranded region includes 2-12 phosphorothioatemodifications. For example, the single stranded region can include 2, 3,4, 5, 6, 7, 8, 9, 10, 11, or 12 phosphorothioate modifications. In someinstances, the single stranded region contains 6-8 phosphorothioatemodifications.

Molecules associated with the invention are also optimized for cellularuptake. In RNA molecules described herein, the guide and/or passengerstrands can be attached to a conjugate. In certain embodiments theconjugate is hydrophobic. The hydrophobic conjugate can be a smallmolecule with a partition coefficient that is higher than 10. Theconjugate can be a sterol-type molecule such as cholesterol, or amolecule with an increased length polycarbon chain attached to C17, andthe presence of a conjugate can influence the ability of an RNA moleculeto be taken into a cell with or without a lipid transfection reagent.The conjugate can be attached to the passenger or guide strand through ahydrophobic linker. In some embodiments, a hydrophobic linker is 5-12Cin length, and/or is hydroxypyrrolidine-based. In some embodiments, ahydrophobic conjugate is attached to the passenger strand and the CUresidues of either the passenger and/or guide strand are modified. Insome embodiments, at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%or 95% of the CU residues on the passenger strand and/or the guidestrand are modified. In some aspects, molecules associated with theinvention are self-delivering (sd). As used herein, “self-delivery”refers to the ability of a molecule to be delivered into a cell withoutthe need for an additional delivery vehicle such as a transfectionreagent.

Aspects of the invention relate to selecting molecules for use in RNAi.Molecules that have a double stranded region of 8-15 nucleotides can beselected for use in RNAi. In some embodiments, molecules are selectedbased on their thermodynamic stability (ΔG). In some embodiments,molecules will be selected that have a (ΔG) of less than −13 kkal/mol.For example, the (ΔG) value may be −13, −14, −15, −16, −17, −18, −19,−21, −22 or less than −22 kkal/mol. In other embodiments, the (ΔG) valuemay be higher than −13 kkal/mol. For example, the (ΔG) value may be −12,−11, −10, −9, −8, −7 or more than −7 kkal/mol. It should be appreciatedthat ΔG can be calculated using any method known in the art. In someembodiments ΔG is calculated using Mfold, available through the Mfoldinternet site (http://mfold.bioinfo.rpi.edu/cgi-bin/rna-form1.cgi)).Methods for calculating ΔG are described in, and are incorporated byreference from, the following references: Zuker, M. (2003) Nucleic AcidsRes., 31(13):3406-15; Mathews, D. H., Sabina, J., Zuker, M. and Turner,D. H. (1999) J. Mol. Biol. 288:911-940; Mathews, D. H., Disney, M. D.,Childs, J. L., Schroeder, S. J., Zuker, M., and Turner, D. H. (2004)Proc. Natl. Acad. Sci. 101:7287-7292; Duan, S., Mathews, D. H., andTurner, D. H. (2006) Biochemistry 45:9819-9832; Wuchty, S., Fontana, W.,Hofacker, I. L., and Schuster, P. (1999) Biopolymers 49:145-165.

In certain embodiments, the polynucleotide contains 5′- and/or 3′-endoverhangs. The number and/or sequence of nucleotides overhang on one endof the polynucleotide may be the same or different from the other end ofthe polynucleotide. In certain embodiments, one or more of the overhangnucleotides may contain chemical modification(s), such asphosphorothioate or 2′-OMe modification.

In certain embodiments, the polynucleotide is unmodified. In otherembodiments, at least one nucleotide is modified. In furtherembodiments, the modification includes a 2′-H or 2′-modified ribosesugar at the 2nd nucleotide from the 5′-end of the guide sequence. The“2nd nucleotide” is defined as the second nucleotide from the 5′-end ofthe polynucleotide.

As used herein, “2′-modified ribose sugar” includes those ribose sugarsthat do not have a 2′-OH group. “2′-modified ribose sugar” does notinclude 2′-deoxyribose (found in unmodified canonical DNA nucleotides).For example, the 2′-modified ribose sugar may be 2′-O-alkyl nucleotides,2′-deoxy-2′-fluoro nucleotides, 2′-deoxy nucleotides, or combinationthereof.

In certain embodiments, the 2′-modified nucleotides are pyrimidinenucleotides (e.g., C/U). Examples of 2′-O-alkyl nucleotides include2′-O-methyl nucleotides, or 2′-O-allyl nucleotides.

In certain embodiments, the sd-rxRNA polynucleotide of the inventionwith the above-referenced 5′-end modification exhibits significantly(e.g., at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90% or more) less “off-target” gene silencing whencompared to similar constructs without the specified 5′-endmodification, thus greatly improving the overall specificity of the RNAireagent or therapeutics.

As used herein, “off-target” gene silencing refers to unintended genesilencing due to, for example, spurious sequence homology between theantisense (guide) sequence and the unintended target mRNA sequence.

According to this aspect of the invention, certain guide strandmodifications further increase nuclease stability, and/or lowerinterferon induction, without significantly decreasing RNAi activity (orno decrease in RNAi activity at all).

In some embodiments, wherein the RNAi construct involves a hairpin, the5′-stem sequence may comprise a 2′-modified ribose sugar, such as2′-O-methyl modified nucleotide, at the 2^(nd) nucleotide on the 5′-endof the polynucleotide and, in some embodiments, no other modifiednucleotides. The hairpin structure having such modification may haveenhanced target specificity or reduced off-target silencing compared toa similar construct without the 2′-O-methyl modification at saidposition.

Certain combinations of specific 5′-stem sequence and 3′-stem sequencemodifications may result in further unexpected advantages, as partlymanifested by enhanced ability to inhibit target gene expression,enhanced serum stability, and/or increased target specificity, etc.

In certain embodiments, the guide strand comprises a 2′-O-methylmodified nucleotide at the 2^(nd) nucleotide on the 5′-end of the guidestrand and no other modified nucleotides.

In other aspects, the sd-rxRNA structures of the present inventionmediates sequence-dependent gene silencing by a microRNA mechanism. Asused herein, the term “microRNA” (“miRNA”), also referred to in the artas “small temporal RNAs” (“stRNAs”), refers to a small (10-50nucleotide) RNA which are genetically encoded (e.g., by viral,mammalian, or plant genomes) and are capable of directing or mediatingRNA silencing. An “miRNA disorder” shall refer to a disease or disordercharacterized by an aberrant expression or activity of an miRNA.

microRNAs are involved in down-regulating target genes in criticalpathways, such as development and cancer, in mice, worms and mammals.Gene silencing through a microRNA mechanism is achieved by specific yetimperfect base-pairing of the miRNA and its target messenger RNA (mRNA).Various mechanisms may be used in microRNA-mediated down-regulation oftarget mRNA expression.

miRNAs are noncoding RNAs of approximately 22 nucleotides which canregulate gene expression at the post transcriptional or translationallevel during plant and animal development. One common feature of miRNAsis that they are all excised from an approximately 70 nucleotideprecursor RNA stem-loop termed pre-miRNA, probably by Dicer, an RNaseIII-type enzyme, or a homolog thereof. Naturally-occurring miRNAs areexpressed by endogenous genes in vivo and are processed from a hairpinor stem-loop precursor (pre-miRNA or pri-miRNAs) by Dicer or otherRNAses. miRNAs can exist transiently in vivo as a double-stranded duplexbut only one strand is taken up by the RISC complex to direct genesilencing.

In some embodiments a version of sd-rxRNA compounds, which are effectivein cellular uptake and inhibiting of miRNA activity are described.Essentially the compounds are similar to RISC entering version but largestrand chemical modification patterns are optimized in the way to blockcleavage and act as an effective inhibitor of the RISC action. Forexample, the compound might be completely or mostly Omethyl modifiedwith the PS content described previously. For these types of compoundsthe 5′ phosphorilation is not necessary. The presence of double strandedregion is preferred as it is promotes cellular uptake and efficient RISCloading.

Another pathway that uses small RNAs as sequence-specific regulators isthe RNA interference (RNAi) pathway, which is an evolutionarilyconserved response to the presence of double-stranded RNA (dsRNA) in thecell. The dsRNAs are cleaved into ˜20-base pair (bp) duplexes ofsmall-interfering RNAs (siRNAs) by Dicer. These small RNAs get assembledinto multiprotein effector complexes called RNA-induced silencingcomplexes (RISCs). The siRNAs then guide the cleavage of target mRNAswith perfect complementarity.

Some aspects of biogenesis, protein complexes, and function are sharedbetween the siRNA pathway and the miRNA pathway. The subjectsingle-stranded polynucleotides may mimic the dsRNA in the siRNAmechanism, or the microRNA in the miRNA mechanism.

In certain embodiments, the modified RNAi constructs may have improvedstability in serum and/or cerebral spinal fluid compared to anunmodified RNAi constructs having the same sequence.

In certain embodiments, the structure of the RNAi construct does notinduce interferon response in primary cells, such as mammalian primarycells, including primary cells from human, mouse and other rodents, andother non-human mammals. In certain embodiments, the RNAi construct mayalso be used to inhibit expression of a target gene in an invertebrateorganism.

To further increase the stability of the subject constructs in vivo, the3′-end of the hairpin structure may be blocked by protective group(s).For example, protective groups such as inverted nucleotides, invertedabasic moieties, or amino-end modified nucleotides may be used. Invertednucleotides may comprise an inverted deoxynucleotide. Inverted abasicmoieties may comprise an inverted deoxyabasic moiety, such as a3′,3′-linked or 5′,5′-linked deoxyabasic moiety.

The RNAi constructs of the invention are capable of inhibiting thesynthesis of any target protein encoded by target gene(s). The inventionincludes methods to inhibit expression of a target gene either in a cellin vitro, or in vivo. As such, the RNAi constructs of the invention areuseful for treating a patient with a disease characterized by theoverexpression of a target gene.

The target gene can be endogenous or exogenous (e.g., introduced into acell by a virus or using recombinant DNA technology) to a cell. Suchmethods may include introduction of RNA into a cell in an amountsufficient to inhibit expression of the target gene. By way of example,such an RNA molecule may have a guide strand that is complementary tothe nucleotide sequence of the target gene, such that the compositioninhibits expression of the target gene.

The invention also relates to vectors expressing the subject hairpinconstructs, and cells comprising such vectors or the subject hairpinconstructs. The cell may be a mammalian cell in vivo or in culture, suchas a human cell.

The invention further relates to compositions comprising the subjectRNAi constructs, and a pharmaceutically acceptable carrier or diluent.

Another aspect of the invention provides a method for inhibiting theexpression of a target gene in a mammalian cell, comprising contactingthe mammalian cell with any of the subject RNAi constructs.

The method may be carried out in vitro, ex vivo, or in vivo, in, forexample, mammalian cells in culture, such as a human cell in culture.

The target cells (e.g., mammalian cell) may be contacted in the presenceof a delivery reagent, such as a lipid (e.g., a cationic lipid) or aliposome.

Another aspect of the invention provides a method for inhibiting theexpression of a target gene in a mammalian cell, comprising contactingthe mammalian cell with a vector expressing the subject RNAi constructs.

In one aspect of the invention, a longer duplex polynucleotide isprovided, including a first polynucleotide that ranges in size fromabout 16 to about 30 nucleotides; a second polynucleotide that ranges insize from about 26 to about 46 nucleotides, wherein the firstpolynucleotide (the antisense strand) is complementary to both thesecond polynucleotide (the sense strand) and a target gene, and whereinboth polynucleotides form a duplex and wherein the first polynucleotidecontains a single stranded region longer than 6 bases in length and ismodified with alternative chemical modification pattern, and/or includesa conjugate moiety that facilitates cellular delivery. In thisembodiment, between about 40% to about 90% of the nucleotides of thepassenger strand between about 40% to about 90% of the nucleotides ofthe guide strand, and between about 40% to about 90% of the nucleotidesof the single stranded region of the first polynucleotide are chemicallymodified nucleotides.

In an embodiment, the chemically modified nucleotide in thepolynucleotide duplex may be any chemically modified nucleotide known inthe art, such as those discussed in detail above. In a particularembodiment, the chemically modified nucleotide is selected from thegroup consisting of 2′ F modified nucleotides, 2′-O-methyl modified and2′deoxy nucleotides. In another particular embodiment, the chemicallymodified nucleotides results from “hydrophobic modifications” of thenucleotide base. In another particular embodiment, the chemicallymodified nucleotides are phosphorothioates. In an additional particularembodiment, chemically modified nucleotides are combination ofphosphorothioates, 2′-O-methyl, 2′deoxy, hydrophobic modifications andphosphorothioates. As these groups of modifications refer tomodification of the ribose ring, back bone and nucleotide, it isfeasible that some modified nucleotides will carry a combination of allthree modification types.

In another embodiment, the chemical modification is not the same acrossthe various regions of the duplex. In a particular embodiment, the firstpolynucleotide (the passenger strand), has a large number of diversechemical modifications in various positions. For this polynucleotide upto 90% of nucleotides might be chemically modified and/or havemismatches introduced. In another embodiment, chemical modifications ofthe first or second polynucleotide include, but not limited to, 5′position modification of Uridine and Cytosine (4-pyridyl, 2-pyridyl,indolyl, phenyl (C₆H₅OH); tryptophanyl (C8H6N)CH2CH(NH2)CO), isobutyl,butyl, aminobenzyl; phenyl; naphthyl, etc), where the chemicalmodification might alter base pairing capabilities of a nucleotide. Forthe guide strand an important feature of this aspect of the invention isthe position of the chemical modification relative to the 5′ end of theantisense and sequence. For example, chemical phosphorylation of the 5′end of the guide strand is usually beneficial for efficacy. O-methylmodifications in the seed region of the sense strand (position 2-7relative to the 5′ end) are not generally well tolerated, whereas 2′Fand deoxy are well tolerated. The mid part of the guide strand and the3′ end of the guide strand are more permissive in a type of chemicalmodifications applied. Deoxy modifications are not tolerated at the 3′end of the guide strand.

A unique feature of this aspect of the invention involves the use ofhydrophobic modification on the bases. In one embodiment, thehydrophobic modifications are preferably positioned near the 5′ end ofthe guide strand, in other embodiments, they localized in the middle ofthe guides strand, in other embodiment they localized at the 3′ end ofthe guide strand and yet in another embodiment they are distributedthought the whole length of the polynucleotide. The same type ofpatterns is applicable to the passenger strand of the duplex.

The other part of the molecule is a single stranded region. The singlestranded region is expected to range from 6 to 40 nucleotides.

In one embodiment, the single stranded region of the firstpolynucleotide contains modifications selected from the group consistingof between 40% and 90% hydrophobic base modifications, between 40%-90%phosphorothioates, between 40%-90% modification of the ribose moiety,and any combination of the preceding.

Efficiency of guide strand (first polynucleotide) loading into the RISCcomplex might be altered for heavily modified polynucleotides, so in oneembodiment, the duplex polynucleotide includes a mismatch betweennucleotide 9, 11, 12, 13, or 14 on the guide strand (firstpolynucleotide) and the opposite nucleotide on the sense strand (secondpolynucleotide) to promote efficient guide strand loading.

More detailed aspects of the invention are described in the sectionsbelow.

Duplex Characteristics

Double-stranded oligonucleotides of the invention may be formed by twoseparate complementary nucleic acid strands. Duplex formation can occureither inside or outside the cell containing the target gene.

As used herein, the term “duplex” includes the region of thedouble-stranded nucleic acid molecule(s) that is (are) hydrogen bondedto a complementary sequence. Double-stranded oligonucleotides of theinvention may comprise a nucleotide sequence that is sense to a targetgene and a complementary sequence that is antisense to the target gene.The sense and antisense nucleotide sequences correspond to the targetgene sequence, e.g., are identical or are sufficiently identical toeffect target gene inhibition (e.g., are about at least about 98%identical, 96% identical, 94%, 90% identical, 85% identical, or 80%identical) to the target gene sequence.

In certain embodiments, the double-stranded oligonucleotide of theinvention is double-stranded over its entire length, i.e., with nooverhanging single-stranded sequence at either end of the molecule,i.e., is blunt-ended. In other embodiments, the individual nucleic acidmolecules can be of different lengths. In other words, a double-strandedoligonucleotide of the invention is not double-stranded over its entirelength. For instance, when two separate nucleic acid molecules are used,one of the molecules, e.g., the first molecule comprising an antisensesequence, can be longer than the second molecule hybridizing thereto(leaving a portion of the molecule single-stranded). Likewise, when asingle nucleic acid molecule is used a portion of the molecule at eitherend can remain single-stranded.

In one embodiment, a double-stranded oligonucleotide of the inventioncontains mismatches and/or loops or bulges, but is double-stranded overat least about 70% of the length of the oligonucleotide. In anotherembodiment, a double-stranded oligonucleotide of the invention isdouble-stranded over at least about 80% of the length of theoligonucleotide. In another embodiment, a double-strandedoligonucleotide of the invention is double-stranded over at least about90%-95% of the length of the oligonucleotide. In another embodiment, adouble-stranded oligonucleotide of the invention is double-stranded overat least about 96%-98% of the length of the oligonucleotide. In certainembodiments, the double-stranded oligonucleotide of the inventioncontains at least or up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, or 15 mismatches.

Modifications

The nucleotides of the invention may be modified at various locations,including the sugar moiety, the phosphodiester linkage, and/or the base.

In some embodiments, the base moiety of a nucleoside may be modified.For example, a pyrimidine base may be modified at the 2, 3, 4, 5, and/or6 position of the pyrimidine ring. In some embodiments, the exocyclicamine of cytosine may be modified. A purine base may also be modified.For example, a purine base may be modified at the 1, 2, 3, 6, 7, or 8position. In some embodiments, the exocyclic amine of adenine may bemodified. In some cases, a nitrogen atom in a ring of a base moiety maybe substituted with another atom, such as carbon. A modification to abase moiety may be any suitable modification. Examples of modificationsare known to those of ordinary skill in the art. In some embodiments,the base modifications include alkylated purines or pyrimidines,acylated purines or pyrimidines, or other heterocycles.

In some embodiments, a pyrimidine may be modified at the 5 position. Forexample, the 5 position of a pyrimidine may be modified with an alkylgroup, an alkynyl group, an alkenyl group, an acyl group, or substitutedderivatives thereof. In other examples, the 5 position of a pyrimidinemay be modified with a hydroxyl group or an alkoxyl group or substitutedderivative thereof. Also, the N⁴ position of a pyrimidine may bealkylated. In still further examples, the pyrimidine 5-6 bond may besaturated, a nitrogen atom within the pyrimidine ring may be substitutedwith a carbon atom, and/or the O² and O⁴ atoms may be substituted withsulfur atoms. It should be understood that other modifications arepossible as well.

In other examples, the N⁷ position and/or N² and/or N³ position of apurine may be modified with an alkyl group or substituted derivativethereof. In further examples, a third ring may be fused to the purinebicyclic ring system and/or a nitrogen atom within the purine ringsystem may be substituted with a carbon atom. It should be understoodthat other modifications are possible as well.

Non-limiting examples of pyrimidines modified at the 5 position aredisclosed in U.S. Pat. No. 5,591,843, U.S. Pat. No. 7,205,297, U.S. Pat.No. 6,432,963, and U.S. Pat. No. 6,020,483; non-limiting examples ofpyrimidines modified at the N⁴ position are disclosed in U.S. Pat. No.5,580,731; non-limiting examples of purines modified at the 8 positionare disclosed in U.S. Pat. No. 6,355,787 and U.S. Pat. No. 5,580,972;non-limiting examples of purines modified at the N⁶ position aredisclosed in U.S. Pat. No. 4,853,386, U.S. Pat. No. 5,789,416, and U.S.Pat. No. 7,041,824; and non-limiting examples of purines modified at the2 position are disclosed in U.S. Pat. No. 4,201,860 and U.S. Pat. No.5,587,469, all of which are incorporated herein by reference.

Non-limiting examples of modified bases include N⁴,N⁴-ethanocytosine,7-deazaxanthosine, 7-deazaguanosine, 8-oxo-N⁶-methyladenine,4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-fluorouracil,5-bromouracil, 5-carboxymethylaminomethyl-2-thiouracil,5-carboxymethylaminomethyl uracil, dihydrouracil, inosine,N⁶-isopentenyl-adenine, 1-methyladenine, 1-methylpseudouracil,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N⁶-methyladenine,7-methylguanine, 5-methylaminomethyl uracil, 5-methoxyaminomethyl-2-thiouracil, 5-methoxyuracil,2-methylthio-N⁶-isopentenyladenine, pseudouracil, 5-methyl-2-thiouracil,2-thiouracil, 4-thiouracil, 5-methyluracil, 2-thiocytosine, and2,6-diaminopurine. In some embodiments, the base moiety may be aheterocyclic base other than a purine or pyrimidine. The heterocyclicbase may be optionally modified and/or substituted.

Sugar moieties include natural, unmodified sugars, e.g., monosaccharide(such as pentose, e.g., ribose, deoxyribose), modified sugars and sugaranalogs. In general, possible modifications of nucleomonomers,particularly of a sugar moiety, include, for example, replacement of oneor more of the hydroxyl groups with a halogen, a heteroatom, analiphatic group, or the functionalization of the hydroxyl group as anether, an amine, a thiol, or the like.

One particularly useful group of modified nucleomonomers are 2′-O-methylnucleotides. Such 2′-O-methyl nucleotides may be referred to as“methylated,” and the corresponding nucleotides may be made fromunmethylated nucleotides followed by alkylation or directly frommethylated nucleotide reagents. Modified nucleomonomers may be used incombination with unmodified nucleomonomers. For example, anoligonucleotide of the invention may contain both methylated andunmethylated nucleomonomers.

Some exemplary modified nucleomonomers include sugar- orbackbone-modified ribonucleotides. Modified ribonucleotides may containa non-naturally occurring base (instead of a naturally occurring base),such as uridines or cytidines modified at the 5′-position, e.g.,5′-(2-amino)propyl uridine and 5′-bromo uridine; adenosines andguanosines modified at the 8-position, e.g., 8-bromo guanosine; deazanucleotides, e.g., 7-deaza-adenosine; and N-alkylated nucleotides, e.g.,N6-methyl adenosine. Also, sugar-modified ribonucleotides may have the2′-OH group replaced by a H, alxoxy (or OR), R or alkyl, halogen, SH,SR, amino (such as NH₂, NHR, NR₂), or CN group, wherein R is loweralkyl, alkenyl, or alkynyl.

Modified ribonucleotides may also have the phosphodiester groupconnecting to adjacent ribonucleotides replaced by a modified group,e.g., of phosphorothioate group. More generally, the various nucleotidemodifications may be combined.

Although the antisense (guide) strand may be substantially identical toat least a portion of the target gene (or genes), at least with respectto the base pairing properties, the sequence need not be perfectlyidentical to be useful, e.g., to inhibit expression of a target gene'sphenotype. Generally, higher homology can be used to compensate for theuse of a shorter antisense gene. In some cases, the antisense strandgenerally will be substantially identical (although in antisenseorientation) to the target gene.

The use of 2′-O-methyl modified RNA may also be beneficial incircumstances in which it is desirable to minimize cellular stressresponses. RNA having 2′-O-methyl nucleomonomers may not be recognizedby cellular machinery that is thought to recognize unmodified RNA. Theuse of 2′-O-methylated or partially 2′-O-methylated RNA may avoid theinterferon response to double-stranded nucleic acids, while maintainingtarget RNA inhibition. This may be useful, for example, for avoiding theinterferon or other cellular stress responses, both in short RNAi (e.g.,siRNA) sequences that induce the interferon response, and in longer RNAisequences that may induce the interferon response.

Overall, modified sugars may include D-ribose, 2′-O-alkyl (including2′-O-methyl and 2′-O-ethyl), i.e., 2′-alkoxy, 2′-amino, 2′-S-alkyl,2′-halo (including 2′-fluoro), 2′-methoxyethoxy, 2′-allyloxy(—OCH₂CH═CH₂), 2′-propargyl, 2′-propyl, ethynyl, ethenyl, propenyl, andcyano and the like. In one embodiment, the sugar moiety can be a hexoseand incorporated into an oligonucleotide as described (Augustyns, K., etal., Nucl. Acids. Res. 18:4711 (1992)). Exemplary nucleomonomers can befound, e.g., in U.S. Pat. No. 5,849,902, incorporated by referenceherein.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. For purposes of this invention, thechemical elements are identified in accordance with the Periodic Tableof the Elements, CAS version, Handbook of Chemistry and Physics, 75^(th)Ed., inside cover, and specific functional groups are generally definedas described therein. Additionally, general principles of organicchemistry, as well as specific functional moieties and reactivity, aredescribed in Organic Chemistry, Thomas Sorrell, University ScienceBooks, Sausalito: 1999, the entire contents of which are incorporatedherein by reference.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, .2 D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

Isomeric mixtures containing any of a variety of isomer ratios may beutilized in accordance with the present invention. For example, whereonly two isomers are combined, mixtures containing 50:50, 60:40, 70:30,80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 isomer ratios areall contemplated by the present invention. Those of ordinary skill inthe art will readily appreciate that analogous ratios are contemplatedfor more complex isomer mixtures.

If, for instance, a particular enantiomer of a compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

In certain embodiments, oligonucleotides of the invention comprise 3′and 5′ termini (except for circular oligonucleotides). In oneembodiment, the 3′ and 5′ termini of an oligonucleotide can besubstantially protected from nucleases e.g., by modifying the 3′ or 5′linkages (e.g., U.S. Pat. No. 5,849,902 and WO 98/13526). For example,oligonucleotides can be made resistant by the inclusion of a “blockinggroup.” The term “blocking group” as used herein refers to substituents(e.g., other than OH groups) that can be attached to oligonucleotides ornucleomonomers, either as protecting groups or coupling groups forsynthesis (e.g., FITC, propyl (CH₂—CH₂—CH₃), glycol (—O—CH₂—CH₂—O—)phosphate (PO₃ ²⁻), hydrogen phosphonate, or phosphoramidite). “Blockinggroups” also include “end blocking groups” or “exonuclease blockinggroups” which protect the 5′ and 3′ termini of the oligonucleotide,including modified nucleotides and non-nucleotide exonuclease resistantstructures.

Exemplary end-blocking groups include cap structures (e.g., a7-methylguanosine cap), inverted nucleomonomers, e.g., with 3′-3′ or5′-5′ end inversions (see, e.g., Ortiagao et al. 1992. Antisense Res.Dev. 2:129), methylphosphonate, phosphoramidite, non-nucleotide groups(e.g., non-nucleotide linkers, amino linkers, conjugates) and the like.The 3′ terminal nucleomonomer can comprise a modified sugar moiety. The3′ terminal nucleomonomer comprises a 3′-0 that can optionally besubstituted by a blocking group that prevents 3′-exonuclease degradationof the oligonucleotide. For example, the 3′-hydroxyl can be esterifiedto a nucleotide through a 3′→3′ internucleotide linkage. For example,the alkyloxy radical can be methoxy, ethoxy, or isopropoxy, andpreferably, ethoxy. Optionally, the 3′→3′linked nucleotide at the 3′terminus can be linked by a substitute linkage. To reduce nucleasedegradation, the 5′ most 3′→5′ linkage can be a modified linkage, e.g.,a phosphorothioate or a P-alkyloxyphosphotriester linkage. Preferably,the two 5′ most 3′→5′ linkages are modified linkages. Optionally, the 5′terminal hydroxy moiety can be esterified with a phosphorus containingmoiety, e.g., phosphate, phosphorothioate, or P-ethoxyphosphate.

One of ordinary skill in the art will appreciate that the syntheticmethods, as described herein, utilize a variety of protecting groups. Bythe term “protecting group,” as used herein, it is meant that aparticular functional moiety, e.g., O, S, or N, is temporarily blockedso that a reaction can be carried out selectively at another reactivesite in a multifunctional compound. In certain embodiments, a protectinggroup reacts selectively in good yield to give a protected substratethat is stable to the projected reactions; the protecting group shouldbe selectively removable in good yield by readily available, preferablynon-toxic reagents that do not attack the other functional groups; theprotecting group forms an easily separable derivative (more preferablywithout the generation of new stereogenic centers); and the protectinggroup has a minimum of additional functionality to avoid further sitesof reaction. As detailed herein, oxygen, sulfur, nitrogen, and carbonprotecting groups may be utilized. Hydroxyl protecting groups includemethyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate(TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutylcarbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts). For protecting 1,2- or 1,3-diols, the protecting groups includemethylene acetal, ethylidene acetal, 1-t-butylethylidene ketal,1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal,2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal,cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal,p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal,3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal,methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethyleneortho ester, 1-methoxyethylidene ortho ester, 1-ethoxyethylidine orthoester, 1,2-dimethoxyethylidene ortho ester, α-methoxybenzylidene orthoester, 1-(N,N-dimethylamino)ethylidene derivative,α-(N,N′-dimethylamino)benzylidene derivative, 2-oxacyclopentylideneortho ester, di-t-butylsilylene group (DTBS),1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative (TIPDS),tetra-t-butoxydisiloxane-1,3-diylidene derivative (TBDS), cycliccarbonates, cyclic boronates, ethyl boronate, and phenyl boronate.Amino-protecting groups include methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, phenothiazinyl-(10)-carbonyl derivative,N′-p-toluenesulfonylaminocarbonyl derivative, N′-phenylaminothiocarbonylderivative, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxycarbonylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate,formamide, acetamide, chloroacetamide, trichloroacetamide,trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide,3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide,acetoacetamide, (N′-dithiobenzyloxycarbonylamino)acetamide,3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethioninederivative, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copperchelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys),p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.Exemplary protecting groups are detailed herein. However, it will beappreciated that the present invention is not intended to be limited tothese protecting groups; rather, a variety of additional equivalentprotecting groups can be readily identified using the above criteria andutilized in the method of the present invention. Additionally, a varietyof protecting groups are described in Protective Groups in OrganicSynthesis, Third Ed. Greene, T. W. and Wuts, P. G., Eds., John Wiley &Sons, New York: 1999, the entire contents of which are herebyincorporated by reference.

It will be appreciated that the compounds, as described herein, may besubstituted with any number of substituents or functional moieties. Ingeneral, the term “substituted” whether preceded by the term“optionally” or not, and substituents contained in formulas of thisinvention, refer to the replacement of hydrogen radicals in a givenstructure with the radical of a specified substituent. When more thanone position in any given structure may be substituted with more thanone substituent selected from a specified group, the substituent may beeither the same or different at every position. As used herein, the term“substituted” is contemplated to include all permissible substituents oforganic compounds. In a broad aspect, the permissible substituentsinclude acyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic substituents of organiccompounds. Heteroatoms such as nitrogen may have hydrogen substituentsand/or any permissible substituents of organic compounds describedherein which satisfy the valencies of the heteroatoms. Furthermore, thisinvention is not intended to be limited in any manner by the permissiblesubstituents of organic compounds. Combinations of substituents andvariables envisioned by this invention are preferably those that resultin the formation of stable compounds useful in the treatment, forexample, of infectious diseases or proliferative disorders. The term“stable”, as used herein, preferably refers to compounds which possessstability sufficient to allow manufacture and which maintain theintegrity of the compound for a sufficient period of time to be detectedand preferably for a sufficient period of time to be useful for thepurposes detailed herein.

The term “aliphatic,” as used herein, includes both saturated andunsaturated, straight chain (i.e., unbranched), branched, acyclic,cyclic, or polycyclic aliphatic hydrocarbons, which are optionallysubstituted with one or more functional groups. As will be appreciatedby one of ordinary skill in the art, “aliphatic” is intended herein toinclude, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, and cycloalkynyl moieties. Thus, as used herein, the term“alkyl” includes straight, branched and cyclic alkyl groups. Ananalogous convention applies to other generic terms such as “alkenyl,”“alkynyl,” and the like. Furthermore, as used herein, the terms “alkyl,”“alkenyl,” “alkynyl,” and the like encompass both substituted andunsubstituted groups. In certain embodiments, as used herein, “loweralkyl” is used to indicate those alkyl groups (cyclic, acyclic,substituted, unsubstituted, branched, or unbranched) having 1-6 carbonatoms.

In certain embodiments, the alkyl, alkenyl, and alkynyl groups employedin the invention contain 1-20 aliphatic carbon atoms. In certain otherembodiments, the alkyl, alkenyl, and alkynyl groups employed in theinvention contain 1-10 aliphatic carbon atoms. In yet other embodiments,the alkyl, alkenyl, and alkynyl groups employed in the invention contain1-8 aliphatic carbon atoms. In still other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-6aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl,and alkynyl groups employed in the invention contain 1-4 carbon atoms.Illustrative aliphatic groups thus include, but are not limited to, forexample, methyl, ethyl, n-propyl, isopropyl, cyclopropyl,—CH₂-cyclopropyl, vinyl, allyl, n-butyl, sec-butyl, isobutyl,tert-butyl, cyclobutyl, —CH₂-cyclobutyl, n-pentyl, sec-pentyl,isopentyl, tert-pentyl, cyclopentyl, —CH₂-cyclopentyl, n-hexyl,sec-hexyl, cyclohexyl, —CH₂-cyclohexyl moieties and the like, whichagain, may bear one or more substituents. Alkenyl groups include, butare not limited to, for example, ethenyl, propenyl, butenyl,1-methyl-2-buten-1-yl, and the like. Representative alkynyl groupsinclude, but are not limited to, ethynyl, 2-propynyl (propargyl),1-propynyl, and the like.

Some examples of substituents of the above-described aliphatic (andother) moieties of compounds of the invention include, but are notlimited to aliphatic; heteroaliphatic; aryl; heteroaryl; arylalkyl;heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; —F; —Cl; —Br; —I;—OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, orheteroarylalkyl, wherein any of the aliphatic, heteroaliphatic,arylalkyl, or heteroarylalkyl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments described herein.

The term “heteroaliphatic,” as used herein, refers to aliphatic moietiesthat contain one or more oxygen, sulfur, nitrogen, phosphorus, orsilicon atoms, e.g., in place of carbon atoms. Heteroaliphatic moietiesmay be branched, unbranched, cyclic or acyclic and include saturated andunsaturated heterocycles such as morpholino, pyrrolidinyl, etc. Incertain embodiments, heteroaliphatic moieties are substituted byindependent replacement of one or more of the hydrogen atoms thereonwith one or more moieties including, but not limited to aliphatic;heteroaliphatic; aryl; heteroaryl; arylalkyl; heteroarylalkyl; alkoxy;aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio;heteroalkylthio; heteroarylthio; —F; —Cl; —Br; —I; —OH; —NO₂; —CN; —CF₃;—CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x);—CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x); —OCO₂R_(x); —OCON(R_(x))₂;—N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x), wherein each occurrence ofR_(x) independently includes, but is not limited to, aliphatic,heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl,wherein any of the aliphatic, heteroaliphatic, arylalkyl, orheteroarylalkyl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substitutents are illustrated by the specificembodiments described herein.

The terms “halo” and “halogen” as used herein refer to an atom selectedfrom fluorine, chlorine, bromine, and iodine.

The term “alkyl” includes saturated aliphatic groups, includingstraight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, nonyl, decyl, etc.), branched-chain alkyl groups(isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl (alicyclic) groups(cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkylsubstituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.In certain embodiments, a straight chain or branched chain alkyl has 6or fewer carbon atoms in its backbone (e.g., C₁-C₆ for straight chain,C₃-C₆ for branched chain), and more preferably 4 or fewer. Likewise,preferred cycloalkyls have from 3-8 carbon atoms in their ringstructure, and more preferably have 5 or 6 carbons in the ringstructure. The term C₁-C₆ includes alkyl groups containing 1 to 6 carbonatoms.

Moreover, unless otherwise specified, the term alkyl includes both“unsubstituted alkyls” and “substituted alkyls,” the latter of whichrefers to alkyl moieties having independently selected substituentsreplacing a hydrogen on one or more carbons of the hydrocarbon backbone.Such substituents can include, for example, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.Cycloalkyls can be further substituted, e.g., with the substituentsdescribed above. An “alkylaryl” or an “arylalkyl” moiety is an alkylsubstituted with an aryl (e.g., phenylmethyl (benzyl)). The term “alkyl”also includes the side chains of natural and unnatural amino acids. Theterm “n-alkyl” means a straight chain (i.e., unbranched) unsubstitutedalkyl group.

The term “alkenyl” includes unsaturated aliphatic groups analogous inlength and possible substitution to the alkyls described above, but thatcontain at least one double bond. For example, the term “alkenyl”includes straight-chain alkenyl groups (e.g., ethylenyl, propenyl,butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.),branched-chain alkenyl groups, cycloalkenyl (alicyclic) groups(cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl,cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl groups, andcycloalkyl or cycloalkenyl substituted alkenyl groups. In certainembodiments, a straight chain or branched chain alkenyl group has 6 orfewer carbon atoms in its backbone (e.g., C₂-C₆ for straight chain,C₃-C₆ for branched chain). Likewise, cycloalkenyl groups may have from3-8 carbon atoms in their ring structure, and more preferably have 5 or6 carbons in the ring structure. The term C₂-C₆ includes alkenyl groupscontaining 2 to 6 carbon atoms.

Moreover, unless otherwise specified, the term alkenyl includes both“unsubstituted alkenyls” and “substituted alkenyls,” the latter of whichrefers to alkenyl moieties having independently selected substituentsreplacing a hydrogen on one or more carbons of the hydrocarbon backbone.Such substituents can include, for example, alkyl groups, alkynylgroups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

The term “alkynyl” includes unsaturated aliphatic groups analogous inlength and possible substitution to the alkyls described above, butwhich contain at least one triple bond. For example, the term “alkynyl”includes straight-chain alkynyl groups (e.g., ethynyl, propynyl,butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.),branched-chain alkynyl groups, and cycloalkyl or cycloalkenylsubstituted alkynyl groups. In certain embodiments, a straight chain orbranched chain alkynyl group has 6 or fewer carbon atoms in its backbone(e.g., C₂-C₆ for straight chain, C₃-C₆ for branched chain). The termC₂-C₆ includes alkynyl groups containing 2 to 6 carbon atoms.

Moreover, unless otherwise specified, the term alkynyl includes both“unsubstituted alkynyls” and “substituted alkynyls,” the latter of whichrefers to alkynyl moieties having independently selected substituentsreplacing a hydrogen on one or more carbons of the hydrocarbon backbone.Such substituents can include, for example, alkyl groups, alkynylgroups, halogens, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto five carbon atoms in its backbone structure. “Lower alkenyl” and“lower alkynyl” have chain lengths of, for example, 2-5 carbon atoms.

The term “alkoxy” includes substituted and unsubstituted alkyl, alkenyl,and alkynyl groups covalently linked to an oxygen atom. Examples ofalkoxy groups include methoxy, ethoxy, isopropyloxy, propoxy, butoxy,and pentoxy groups. Examples of substituted alkoxy groups includehalogenated alkoxy groups. The alkoxy groups can be substituted withindependently selected groups such as alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulffiydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfmyl, sulfonato,sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.Examples of halogen substituted alkoxy groups include, but are notlimited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,chloromethoxy, dichloromethoxy, trichloromethoxy, etc.

The term “hydrophobic modifications’ include bases modified in afashion, where (1) overall hydrophobicity of the base is significantlyincreases, (2) the base is still capable of forming close to regularWatson-Crick interaction. Some, of the examples of base modificationsinclude but are not limited to 5-position uridine and cytidinemodifications like phenyl,

4-pyridyl, 2-pyridyl, indolyl, and isobutyl, phenyl (C6H5OH);tryptophanyl (C8H6N)CH2CH(NH2)CO), Isobutyl, butyl, aminobenzyl; phenyl;naphthyl, For purposes of the present invention, the term “overhang”refers to terminal non-base pairing nucleotide(s) resulting from onestrand or region extending beyond the terminus of the complementarystrand to which the first strand or region forms a duplex. One or morepolynucleotides that are capable of forming a duplex through hydrogenbonding can have overhangs. The overhand length generally doesn't exceed5 bases in length.

The term “heteroatom” includes atoms of any element other than carbon orhydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur andphosphorus.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O⁻(with an appropriate counterion).

The term “halogen” includes fluorine, bromine, chlorine, iodine, etc.The term “perhalogenated” generally refers to a moiety wherein allhydrogens are replaced by halogen atoms.

The term “substituted” includes independently selected substituentswhich can be placed on the moiety and which allow the molecule toperform its intended function. Examples of substituents include alkyl,alkenyl, alkynyl, aryl, (CR′R″)₀₋₃NR′R″, (CR′R″)₀₋₃CN, NO₂, halogen,(CR′R″)₀₋₃C(halogen)₃, (CR′R″)₀₋₃CH(halogen)₂, (CR′R″)₀₋₃CH₂(halogen),(CR′R″)₀₋₃CONR′R″, (CR′R″)₀₋₃S(O)₁₋₂NR′R″, (CR′R″)₀₋₃CHO,(CR′R″)₀₋₃O(CR′R″)₀₋₃H, (CR′R″)₀₋₃S(O)₀₋₂R′, (CR′R″)₀₋₃O(CR′R″)₀₋₃H,(CR′R″)₀₋₃COR′, (CR′R″)₀₋₃CO₂R′, or (CR′R″)₀₋₃OR′ groups; wherein eachR′ and R″ are each independently hydrogen, a C₁-C₅ alkyl, C₂-C₅ alkenyl,C₂-C₅ alkynyl, or aryl group, or R′ and R″ taken together are abenzylidene group or a —(CH₂)₂O(CH₂)₂— group.

The term “amine” or “amino” includes compounds or moieties in which anitrogen atom is covalently bonded to at least one carbon or heteroatom.The term “alkyl amino” includes groups and compounds wherein thenitrogen is bound to at least one additional alkyl group. The term“dialkyl amino” includes groups wherein the nitrogen atom is bound to atleast two additional alkyl groups.

The term “ether” includes compounds or moieties which contain an oxygenbonded to two different carbon atoms or heteroatoms. For example, theterm includes “alkoxyalkyl,” which refers to an alkyl, alkenyl, oralkynyl group covalently bonded to an oxygen atom which is covalentlybonded to another alkyl group.

The terms “polynucleotide,” “nucleotide sequence,” “nucleic acid,”“nucleic acid molecule,” “nucleic acid sequence,” and “oligonucleotide”refer to a polymer of two or more nucleotides. The polynucleotides canbe DNA, RNA, or derivatives or modified versions thereof. Thepolynucleotide may be single-stranded or double-stranded. Thepolynucleotide can be modified at the base moiety, sugar moiety, orphosphate backbone, for example, to improve stability of the molecule,its hybridization parameters, etc. The polynucleotide may comprise amodified base moiety which is selected from the group including but notlimited to 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil,hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil,beta-D-galactosylqueosine, inosine, N6-isopentenyladenine,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine,7-methylguanine, 5-methylaminomethyluracil,5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5′-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N6-isopentenyladenine, wybutoxosine, pseudouracil,queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil,4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester,uracil-5-oxyacetic acid, 5-methyl-2-thiouracil,3-(3-amino-3-N-2-carboxypropyl) uracil, and 2,6-diaminopurine. Thepolynucleotide may comprise a modified sugar moiety (e.g.,2′-fluororibose, ribose, 2′-deoxyribose, 2′-O-methylcytidine, arabinose,and hexose), and/or a modified phosphate moiety (e.g., phosphorothioatesand 5′-N-phosphoramidite linkages). A nucleotide sequence typicallycarries genetic information, including the information used by cellularmachinery to make proteins and enzymes. These terms include double- orsingle-stranded genomic and cDNA, RNA, any synthetic and geneticallymanipulated polynucleotide, and both sense and antisensepolynucleotides. This includes single- and double-stranded molecules,i.e., DNA-DNA, DNA-RNA, and RNA-RNA hybrids, as well as “protein nucleicacids” (PNA) formed by conjugating bases to an amino acid backbone.

The term “base” includes the known purine and pyrimidine heterocyclicbases, deazapurines, and analogs (including heterocyclic substitutedanalogs, e.g., aminoethyoxy phenoxazine), derivatives (e.g., 1-alkyl-,1-alkenyl-, heteroaromatic- and 1-alkynyl derivatives) and tautomersthereof. Examples of purines include adenine, guanine, inosine,diaminopurine, and xanthine and analogs (e.g., 8-oxo-N⁶-methyladenine or7-diazaxanthine) and derivatives thereof. Pyrimidines include, forexample, thymine, uracil, and cytosine, and their analogs (e.g.,5-methylcytosine, 5-methyluracil, 5-(1-propynyl)uracil,5-(1-propynyl)cytosine and 4,4-ethanocytosine). Other examples ofsuitable bases include non-purinyl and non-pyrimidinyl bases such as2-aminopyridine and triazines.

In a preferred embodiment, the nucleomonomers of an oligonucleotide ofthe invention are RNA nucleotides. In another preferred embodiment, thenucleomonomers of an oligonucleotide of the invention are modified RNAnucleotides. Thus, the oligonucleotides contain modified RNAnucleotides.

The term “nucleoside” includes bases which are covalently attached to asugar moiety, preferably ribose or deoxyribose. Examples of preferrednucleosides include ribonucleosides and deoxyribonucleosides.Nucleosides also include bases linked to amino acids or amino acidanalogs which may comprise free carboxyl groups, free amino groups, orprotecting groups. Suitable protecting groups are well known in the art(see P. G. M. Wuts and T. W. Greene, “Protective Groups in OrganicSynthesis”, 2^(nd) Ed., Wiley-Interscience, New York, 1999).

The term “nucleotide” includes nucleosides which further comprise aphosphate group or a phosphate analog.

The nucleic acid molecules may be associated with a hydrophobic moietyfor targeting and/or delivery of the molecule to a cell. In certainembodiments, the hydrophobic moiety is associated with the nucleic acidmolecule through a linker. In certain embodiments, the association isthrough non-covalent interactions. In other embodiments, the associationis through a covalent bond. Any linker known in the art may be used toassociate the nucleic acid with the hydrophobic moiety. Linkers known inthe art are described in published international PCT applications, WO92/03464, WO 95/23162, WO 2008/021157, WO 2009/021157, WO 2009/134487,WO 2009/126933, U.S. Patent Application Publication 2005/0107325, U.S.Pat. No. 5,414,077, U.S. Pat. No. 5,419,966, U.S. Pat. No. 5,512,667,U.S. Pat. No. 5,646,126, and U.S. Pat. No. 5,652,359, which areincorporated herein by reference. The linker may be as simple as acovalent bond to a multi-atom linker. The linker may be cyclic oracyclic. The linker may be optionally substituted. In certainembodiments, the linker is capable of being cleaved from the nucleicacid. In certain embodiments, the linker is capable of being hydrolyzedunder physiological conditions. In certain embodiments, the linker iscapable of being cleaved by an enzyme (e.g., an esterase orphosphodiesterase). In certain embodiments, the linker comprises aspacer element to separate the nucleic acid from the hydrophobic moiety.The spacer element may include one to thirty carbon or heteroatoms. Incertain embodiments, the linker and/or spacer element comprisesprotonatable functional groups. Such protonatable functional groups maypromote the endosomal escape of the nucleic acid molecule. Theprotonatable functional groups may also aid in the delivery of thenucleic acid to a cell, for example, neutralizing the overall charge ofthe molecule. In other embodiments, the linker and/or spacer element isbiologically inert (that is, it does not impart biological activity orfunction to the resulting nucleic acid molecule).

In certain embodiments, the nucleic acid molecule with a linker andhydrophobic moiety is of the formulae described herein. In certainembodiments, the nucleic acid molecule is of the formula:

whereinX is N or CH;A is a bond; substituted or unsubstituted, cyclic or acyclic, branchedor unbranched aliphatic; or substituted or unsubstituted, cyclic oracyclic, branched or unbranched heteroaliphatic;R¹ is a hydrophobic moiety;R² is hydrogen; an oxygen-protecting group; cyclic or acyclic,substituted or unsubstituted, branched or unbranched aliphatic; cyclicor acyclic, substituted or unsubstituted, branched or unbranchedheteroaliphatic; substituted or unsubstituted, branched or unbranchedacyl; substituted or unsubstituted, branched or unbranched aryl;substituted or unsubstituted, branched or unbranched heteroaryl; andR³ is a nucleic acid.In certain embodiments, the molecule is of the formula:

In certain embodiments, the molecule is of the formula:

In certain embodiments, the molecule is of the formula:

In certain embodiments, the molecule is of the formula:

In certain embodiments, X is N. In certain embodiments, X is CH.In certain embodiments, A is a bond. In certain embodiments, A issubstituted or unsubstituted, cyclic or acyclic, branched or unbranchedaliphatic. In certain embodiments, A is acyclic, substituted orunsubstituted, branched or unbranched aliphatic. In certain embodiments,A is acyclic, substituted, branched or unbranched aliphatic. In certainembodiments, A is acyclic, substituted, unbranched aliphatic. In certainembodiments, A is acyclic, substituted, unbranched alkyl. In certainembodiments, A is acyclic, substituted, unbranched C₁₋₂₀ alkyl. Incertain embodiments, A is acyclic, substituted, unbranched C₁₋₁₂ alkyl.In certain embodiments, A is acyclic, substituted, unbranched C₁₋₁₀alkyl. In certain embodiments, A is acyclic, substituted, unbranchedC₁₋₈ alkyl. In certain embodiments, A is acyclic, substituted,unbranched C₁₋₆ alkyl. In certain embodiments, A is substituted orunsubstituted, cyclic or acyclic, branched or unbranchedheteroaliphatic. In certain embodiments, A is acyclic, substituted orunsubstituted, branched or unbranched heteroaliphatic. In certainembodiments, A is acyclic, substituted, branched or unbranchedheteroaliphatic. In certain embodiments, A is acyclic, substituted,unbranched heteroaliphatic.In certain embodiments, A is of the formula:

In certain embodiments, A is of one of the formulae:

In certain embodiments, A is of one of the formulae:

In certain embodiments, A is of one of the formulae:

In certain embodiments, A is of the formula:

In certain embodiments, A is of the formula:

In certain embodiments, A is of the formula:

whereineach occurrence of R is independently the side chain of a natural orunnatural amino acid; andn is an integer between 1 and 20, inclusive. In certain embodiments, Ais of the formula:

In certain embodiments, each occurrence of R is independently the sidechain of a natural amino acid. In certain embodiments, n is an integerbetween 1 and 15, inclusive. In certain embodiments, n is an integerbetween 1 and 10, inclusive. In certain embodiments, n is an integerbetween 1 and 5, inclusive.

In certain embodiments, A is of the formula:

wherein n is an integer between 1 and 20, inclusive. In certainembodiments, A is of the formula:

In certain embodiments, n is an integer between 1 and 15, inclusive. Incertain embodiments, n is an integer between 1 and 10, inclusive. Incertain embodiments, n is an integer between 1 and 5, inclusive.In certain embodiments, A is of the formula:

wherein n is an integer between 1 and 20, inclusive. In certainembodiments, A is of the formula:

In certain embodiments, n is an integer between 1 and 15, inclusive. Incertain embodiments, n is an integer between 1 and 10, inclusive. Incertain embodiments, n is an integer between 1 and 5, inclusive.In certain embodiments, the molecule is of the formula:

wherein X, R¹, R², and R³ are as defined herein; andA′ is substituted or unsubstituted, cyclic or acyclic, branched orunbranched aliphatic; or substituted or unsubstituted, cyclic oracyclic, branched or unbranched heteroaliphatic.In certain embodiments, A′ is of one of the formulae:

In certain embodiments, A is of one of the formulae:

In certain embodiments, A is of one of the formulae:

In certain embodiments, A is of the formula:

In certain embodiments, A is of the formula:

In certain embodiments, R¹ is a steroid. In certain embodiments, R¹ is acholesterol. In certain embodiments, R¹ is a lipophilic vitamin. Incertain embodiments, R¹ is a vitamin A. In certain embodiments, R¹ is avitamin E.In certain embodiments, R¹ is of the formula:

wherein R^(A) is substituted or unsubstituted, cyclic or acyclic,branched or unbranched aliphatic; or substituted or unsubstituted,cyclic or acyclic, branched or unbranched heteroaliphatic.In certain embodiments, R¹ is of the formula:

In certain embodiments, R¹ is of the formula:

In certain embodiments, R¹ is of the formula:

In certain embodiments, R¹ is of the formula:

In certain embodiments, R¹ is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

whereinX is N or CH;A is a bond; substituted or unsubstituted, cyclic or acyclic, branchedor unbranched aliphatic; or substituted or unsubstituted, cyclic oracyclic, branched or unbranched heteroaliphatic;R¹ is a hydrophobic moiety;R² is hydrogen; an oxygen-protecting group; cyclic or acyclic,substituted or unsubstituted, branched or unbranched aliphatic; cyclicor acyclic, substituted or unsubstituted, branched or unbranchedheteroaliphatic; substituted or unsubstituted, branched or unbranchedacyl; substituted or unsubstituted, branched or unbranched aryl;substituted or unsubstituted, branched or unbranched heteroaryl; andR³ is a nucleic acid.In certain embodiments, the nucleic acid molecule is of the formula:

whereinX is N or CH;A is a bond; substituted or unsubstituted, cyclic or acyclic, branchedor unbranched aliphatic; or substituted or unsubstituted, cyclic oracyclic, branched or unbranched heteroaliphatic;R¹ is a hydrophobic moiety;R² is hydrogen; an oxygen-protecting group; cyclic or acyclic,substituted or unsubstituted, branched or unbranched aliphatic; cyclicor acyclic, substituted or unsubstituted, branched or unbranchedheteroaliphatic; substituted or unsubstituted, branched or unbranchedacyl; substituted or unsubstituted, branched or unbranched aryl;substituted or unsubstituted, branched or unbranched heteroaryl; andR³ is a nucleic acid.In certain embodiments, the nucleic acid molecule is of the formula:

whereinX is N or CH;A is a bond; substituted or unsubstituted, cyclic or acyclic, branchedor unbranched aliphatic; or substituted or unsubstituted, cyclic oracyclic, branched or unbranched heteroaliphatic;R¹ is a hydrophobic moiety;R² is hydrogen; an oxygen-protecting group; cyclic or acyclic,substituted or unsubstituted, branched or unbranched aliphatic; cyclicor acyclic, substituted or unsubstituted, branched or unbranchedheteroaliphatic; substituted or unsubstituted, branched or unbranchedacyl; substituted or unsubstituted, branched or unbranched aryl;substituted or unsubstituted, branched or unbranched heteroaryl; andR³ is a nucleic acid. In certain embodiments, the nucleic acid moleculeis of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

wherein R³ is a nucleic acid.In certain embodiments, the nucleic acid molecule is of the formula:

wherein R³ is a nucleic acid; andn is an integer between 1 and 20, inclusive.In certain embodiments, the nucleic acid molecule is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

In certain embodiments, the nucleic acid molecule is of the formula:

As used herein, the term “linkage” includes a naturally occurring,unmodified phosphodiester moiety (—O—(PO²⁻)—O—) that covalently couplesadjacent nucleomonomers. As used herein, the term “substitute linkage”includes any analog or derivative of the native phosphodiester groupthat covalently couples adjacent nucleomonomers. Substitute linkagesinclude phosphodiester analogs, e.g., phosphorothioate,phosphorodithioate, and P-ethyoxyphosphodiester, P-ethoxyphosphodiester,P-alkyloxyphosphotriester, methylphosphonate, and nonphosphoruscontaining linkages, e.g., acetals and amides. Such substitute linkagesare known in the art (e.g., Bjergarde et al. 1991. Nucleic Acids Res.19:5843; Caruthers et al. 1991. Nucleosides Nucleotides. 10:47). Incertain embodiments, non-hydrolizable linkages are preferred, such asphosphorothioate linkages.

In certain embodiments, oligonucleotides of the invention comprisehydrophobicly modified nucleotides or “hydrophobic modifications.” Asused herein “hydrophobic modifications” refers to bases that aremodified such that (1) overall hydrophobicity of the base issignificantly increased, and/or (2) the base is still capable of formingclose to regular Watson-Crick interaction. Several non-limiting examplesof base modifications include 5-position uridine and cytidinemodifications such as phenyl, 4-pyridyl, 2-pyridyl, indolyl, andisobutyl, phenyl (C6H5OH); tryptophanyl (C8H6N)CH2CH(NH2)CO), Isobutyl,butyl, aminobenzyl; phenyl; and naphthyl.

Another type of conjugates that can be attached to the end (3′ or 5′end), the loop region, or any other parts of the sd-rxRNA might includea sterol, sterol type molecule, peptide, small molecule, protein, etc.In some embodiments, a sd-rxRNA may contain more than one conjugates(same or different chemical nature). In some embodiments, the conjugateis cholesterol.

Another way to increase target gene specificity, or to reduce off-targetsilencing effect, is to introduce a 2′-modification (such as the 2′-Omethyl modification) at a position corresponding to the second 5′-endnucleotide of the guide sequence. This allows the positioning of this2′-modification in the Dicer-resistant hairpin structure, thus enablingone to design better RNAi constructs with less or no off-targetsilencing.

In one embodiment, a hairpin polynucleotide of the invention cancomprise one nucleic acid portion which is DNA and one nucleic acidportion which is RNA. Antisense (guide) sequences of the invention canbe “chimeric oligonucleotides” which comprise an RNA-like and a DNA-likeregion.

The language “RNase H activating region” includes a region of anoligonucleotide, e.g., a chimeric oligonucleotide, that is capable ofrecruiting RNase H to cleave the target RNA strand to which theoligonucleotide binds. Typically, the RNase activating region contains aminimal core (of at least about 3-5, typically between about 3-12, moretypically, between about 5-12, and more preferably between about 5-10contiguous nucleomonomers) of DNA or DNA-like nucleomonomers. (See,e.g., U.S. Pat. No. 5,849,902). Preferably, the RNase H activatingregion comprises about nine contiguous deoxyribose containingnucleomonomers.

The language “non-activating region” includes a region of an antisensesequence, e.g., a chimeric oligonucleotide, that does not recruit oractivate RNase H. Preferably, a non-activating region does not comprisephosphorothioate DNA. The oligonucleotides of the invention comprise atleast one non-activating region. In one embodiment, the non-activatingregion can be stabilized against nucleases or can provide specificityfor the target by being complementary to the target and forming hydrogenbonds with the target nucleic acid molecule, which is to be bound by theoligonucleotide.

In one embodiment, at least a portion of the contiguous polynucleotidesare linked by a substitute linkage, e.g., a phosphorothioate linkage.

In certain embodiments, most or all of the nucleotides beyond the guidesequence (2′-modified or not) are linked by phosphorothioate linkages.Such constructs tend to have improved pharmacokinetics due to theirhigher affinity for serum proteins. The phosphorothioate linkages in thenon-guide sequence portion of the polynucleotide generally do notinterfere with guide strand activity, once the latter is loaded intoRISC.

Antisense (guide) sequences of the present invention may include“morpholino oligonucleotides.” Morpholino oligonucleotides are non-ionicand function by an RNase H-independent mechanism. Each of the 4 geneticbases (Adenine, Cytosine, Guanine, and Thymine/Uracil) of the morpholinooligonucleotides is linked to a 6-membered morpholine ring. Morpholinooligonucleotides are made by joining the 4 different subunit types by,e.g., non-ionic phosphorodiamidate inter-subunit linkages. Morpholinooligonucleotides have many advantages including: complete resistance tonucleases (Antisense & Nucl. Acid Drug Dev. 1996. 6:267); predictabletargeting (Biochemica Biophysica Acta. 1999. 1489:141); reliableactivity in cells (Antisense & Nucl. Acid Drug Dev. 1997. 7:63);excellent sequence specificity (Antisense & Nucl. Acid Drug Dev. 1997.7:151); minimal non-antisense activity (Biochemica Biophysica Acta.1999. 1489:141); and simple osmotic or scrape delivery (Antisense &Nucl. Acid Drug Dev. 1997. 7:291). Morpholino oligonucleotides are alsopreferred because of their non-toxicity at high doses. A discussion ofthe preparation of morpholino oligonucleotides can be found in Antisense& Nucl. Acid Drug Dev. 1997. 7:187.

The chemical modifications described herein are believed, based on thedata described herein, to promote single stranded polynucleotide loadinginto the RISC. Single stranded polynucleotides have been shown to beactive in loading into RISC and inducing gene silencing. However, thelevel of activity for single stranded polynucleotides appears to be 2 to4 orders of magnitude lower when compared to a duplex polynucleotide.

The present invention provides a description of the chemicalmodification patterns, which may (a) significantly increase stability ofthe single stranded polynucleotide (b) promote efficient loading of thepolynucleotide into the RISC complex and (c) improve uptake of thesingle stranded nucleotide by the cell. FIG. 5 provides somenon-limiting examples of the chemical modification patterns which may bebeneficial for achieving single stranded polynucleotide efficacy insidethe cell. The chemical modification patterns may include combination ofribose, backbone, hydrophobic nucleoside and conjugate type ofmodifications. In addition, in some of the embodiments, the 5′ end ofthe single polynucleotide may be chemically phosphorylated.

In yet another embodiment, the present invention provides a descriptionof the chemical modifications patterns, which improve functionality ofRISC inhibiting polynucleotides. Single stranded polynucleotides havebeen shown to inhibit activity of a preloaded RISC complex through thesubstrate competition mechanism. For these types of molecules,conventionally called antagomers, the activity usually requires highconcentration and in vivo delivery is not very effective. The presentinvention provides a description of the chemical modification patterns,which may (a) significantly increase stability of the single strandedpolynucleotide (b) promote efficient recognition of the polynucleotideby the RISC as a substrate and/or (c) improve uptake of the singlestranded nucleotide by the cell. FIG. 6 provides some non-limitingexamples of the chemical modification patterns that may be beneficialfor achieving single stranded polynucleotide efficacy inside the cell.The chemical modification patterns may include combination of ribose,backbone, hydrophobic nucleoside and conjugate type of modifications.

The modifications provided by the present invention are applicable toall polynucleotides. This includes single stranded RISC enteringpolynucleotides, single stranded RISC inhibiting polynucleotides,conventional duplexed polynucleotides of variable length (15-40 bp),asymmetric duplexed polynucleotides, and the like. Polynucleotides maybe modified with wide variety of chemical modification patterns,including 5′ end, ribose, backbone and hydrophobic nucleosidemodifications.

Synthesis

Oligonucleotides of the invention can be synthesized by any method knownin the art, e.g., using enzymatic synthesis and/or chemical synthesis.The oligonucleotides can be synthesized in vitro (e.g., using enzymaticsynthesis and chemical synthesis) or in vivo (using recombinant DNAtechnology well known in the art).

In a preferred embodiment, chemical synthesis is used for modifiedpolynucleotides. Chemical synthesis of linear oligonucleotides is wellknown in the art and can be achieved by solution or solid phasetechniques. Preferably, synthesis is by solid phase methods.Oligonucleotides can be made by any of several different syntheticprocedures including the phosphoramidite, phosphite triester,H-phosphonate, and phosphotriester methods, typically by automatedsynthesis methods.

Oligonucleotide synthesis protocols are well known in the art and can befound, e.g., in U.S. Pat. No. 5,830,653; WO 98/13526; Stec et al. 1984.J. Am. Chem. Soc. 106:6077; Stec et al. 1985. J. Org. Chem. 50:3908;Stec et al. J. Chromatog. 1985. 326:263; LaPlanche et al. 1986. Nucl.Acid. Res. 1986. 14:9081; Fasman G. D., 1989. Practical Handbook ofBiochemistry and Molecular Biology. 1989. CRC Press, Boca Raton, Fla.;Lamone. 1993. Biochem. Soc. Trans. 21:1; U.S. Pat. No. 5,013,830; U.S.Pat. No. 5,214,135; U.S. Pat. No. 5,525,719; Kawasaki et al. 1993. J.Med. Chem. 36:831; WO 92/03568; U.S. Pat. No. 5,276,019; and U.S. Pat.No. 5,264,423.

The synthesis method selected can depend on the length of the desiredoligonucleotide and such choice is within the skill of the ordinaryartisan. For example, the phosphoramidite and phosphite triester methodcan produce oligonucleotides having 175 or more nucleotides, while theH-phosphonate method works well for oligonucleotides of less than 100nucleotides. If modified bases are incorporated into theoligonucleotide, and particularly if modified phosphodiester linkagesare used, then the synthetic procedures are altered as needed accordingto known procedures. In this regard, Uhlmann et al. (1990, ChemicalReviews 90:543-584) provide references and outline procedures for makingoligonucleotides with modified bases and modified phosphodiesterlinkages. Other exemplary methods for making oligonucleotides are taughtin Sonveaux. 1994. “Protecting Groups in Oligonucleotide Synthesis”;Agrawal. Methods in Molecular Biology 26:1. Exemplary synthesis methodsare also taught in “Oligonucleotide Synthesis—A Practical Approach”(Gait, M. J. IRL Press at Oxford University Press. 1984). Moreover,linear oligonucleotides of defined sequence, including some sequenceswith modified nucleotides, are readily available from several commercialsources.

The oligonucleotides may be purified by polyacrylamide gelelectrophoresis, or by any of a number of chromatographic methods,including gel chromatography and high pressure liquid chromatography. Toconfirm a nucleotide sequence, especially unmodified nucleotidesequences, oligonucleotides may be subjected to DNA sequencing by any ofthe known procedures, including Maxam and Gilbert sequencing, Sangersequencing, capillary electrophoresis sequencing, the wandering spotsequencing procedure or by using selective chemical degradation ofoligonucleotides bound to Hybond paper. Sequences of shortoligonucleotides can also be analyzed by laser desorption massspectroscopy or by fast atom bombardment (McNeal, et al., 1982, J. Am.Chem. Soc. 104:976; Viari, et al., 1987, Biomed. Environ. Mass Spectrom.14:83; Grotjahn et al., 1982, Nuc. Acid Res. 10:4671). Sequencingmethods are also available for RNA oligonucleotides.

The quality of oligonucleotides synthesized can be verified by testingthe oligonucleotide by capillary electrophoresis and denaturing stronganion HPLC (SAX-HPLC) using, e.g., the method of Bergot and Egan. 1992.J. Chrom. 599:35.

Other exemplary synthesis techniques are well known in the art (see,e.g., Sambrook et al., Molecular Cloning: a Laboratory Manual, SecondEdition (1989); DNA Cloning, Volumes I and II (DN Glover Ed. 1985);Oligonucleotide Synthesis (M J Gait Ed, 1984; Nucleic Acid Hybridisation(B D Hames and S J Higgins eds. 1984); A Practical Guide to MolecularCloning (1984); or the series, Methods in Enzymology (Academic Press,Inc.)).

In certain embodiments, the subject RNAi constructs or at least portionsthereof are transcribed from expression vectors encoding the subjectconstructs. Any art recognized vectors may be use for this purpose. Thetranscribed RNAi constructs may be isolated and purified, before desiredmodifications (such as replacing an unmodified sense strand with amodified one, etc.) are carried out.

Delivery/Carrier

Uptake of Oligonucleotides by Cells

Oligonucleotides and oligonucleotide compositions are contacted with(i.e., brought into contact with, also referred to herein asadministered or delivered to) and taken up by one or more cells or acell lysate. The term “cells” includes prokaryotic and eukaryotic cells,preferably vertebrate cells, and, more preferably, mammalian cells. In apreferred embodiment, the oligonucleotide compositions of the inventionare contacted with human cells.

Oligonucleotide compositions of the invention can be contacted withcells in vitro, e.g., in a test tube or culture dish, (and may or maynot be introduced into a subject) or in vivo, e.g., in a subject such asa mammalian subject. Oligonucleotides are taken up by cells at a slowrate by endocytosis, but endocytosed oligonucleotides are generallysequestered and not available, e.g., for hybridization to a targetnucleic acid molecule. In one embodiment, cellular uptake can befacilitated by electroporation or calcium phosphate precipitation.However, these procedures are only useful for in vitro or ex vivoembodiments, are not convenient and, in some cases, are associated withcell toxicity.

In another embodiment, delivery of oligonucleotides into cells can beenhanced by suitable art recognized methods including calcium phosphate,DMSO, glycerol or dextran, electroporation, or by transfection, e.g.,using cationic, anionic, or neutral lipid compositions or liposomesusing methods known in the art (see e.g., WO 90/14074; WO 91/16024; WO91/17424; U.S. Pat. No. 4,897,355; Bergan et al. 1993. Nucleic AcidsResearch. 21:3567). Enhanced delivery of oligonucleotides can also bemediated by the use of vectors (See e.g., Shi, Y. 2003. Trends Genet2003 Jan. 19:9; Reichhart J Metal. Genesis. 2002. 34(1-2):1604, Yu etal. 2002. Proc. Natl. Acad Sci. USA 99:6047; Sui et al. 2002. Proc.Natl. Acad Sci. USA 99:5515) viruses, polyamine or polycation conjugatesusing compounds such as polylysine, protamine, or Ni, N12-bis (ethyl)spermine (see, e.g., Bartzatt, R. et al. 1989. Biotechnol. Appl.Biochem. 11:133; Wagner E. et al. 1992. Proc. Natl. Acad. Sci. 88:4255).

In certain embodiments, the sd-rxRNA of the invention may be deliveredby using various beta-glucan containing particles, referred to as GeRPs(glucan encapsulated RNA loaded particle), described in, andincorporated by reference from, U.S. Provisional Application No.61/310,611, filed on Mar. 4, 2010 and entitled “Formulations and Methodsfor Targeted Delivery to Phagocyte Cells.” Such particles are alsodescribed in, and incorporated by reference from US Patent PublicationsUS 2005/0281781 A1, and US 2010/0040656, and in PCT publications WO2006/007372, and WO 2007/050643. The sd-rxRNA molecule may behydrophobically modified and optionally may be associated with a lipidand/or amphiphilic peptide. In certain embodiments, the beta-glucanparticle is derived from yeast. In certain embodiments, the payloadtrapping molecule is a polymer, such as those with a molecular weight ofat least about 1000 Da, 10,000 Da, 50,000 Da, 100 kDa, 500 kDa, etc.Preferred polymers include (without limitation) cationic polymers,chitosans, or PEI (polyethylenimine), etc.

Glucan particles can be derived from insoluble components of fungal cellwalls such as yeast cell walls. In some embodiments, the yeast isBaker's yeast. Yeast-derived glucan molecules can include one or more ofβ-(1,3)-Glucan, β-(1,6)-Glucan, mannan and chitin. In some embodiments,a glucan particle comprises a hollow yeast cell wall whereby theparticle maintains a three dimensional structure resembling a cell,within which it can complex with or encapsulate a molecule such as anRNA molecule. Some of the advantages associated with the use of yeastcell wall particles are availability of the components, theirbiodegradable nature, and their ability to be targeted to phagocyticcells.

In some embodiments, glucan particles can be prepared by extraction ofinsoluble components from cell walls, for example by extracting Baker'syeast (Fleischmann's) with 1M NaOH/pH 4.0 H2O, followed by washing anddrying. Methods of preparing yeast cell wall particles are discussed in,and incorporated by reference from U.S. Pat. Nos. 4,810,646, 4,992,540,5,082,936, 5,028,703, 5,032,401, 5,322,841, 5,401,727, 5,504,079,5,607,677, 5,968,811, 6,242,594, 6,444,448, 6,476,003, US PatentPublications 2003/0216346, 2004/0014715 and 2010/0040656, and PCTpublished application WO02/12348.

Protocols for preparing glucan particles are also described in, andincorporated by reference from, the following references: Soto andOstroff (2008), “Characterization of multilayered nanoparticlesencapsulated in yeast cell wall particles for DNA delivery.” BioconjugChem 19(4):840-8; Soto and Ostroff (2007), “Oral Macrophage MediatedGene Delivery System,” Nanotech, Volume 2, Chapter 5 (“Drug Delivery”),pages 378-381; and Li et al. (2007), “Yeast glucan particles activatemurine resident macrophages to secrete proinflammatory cytokines viaMyD88- and Syk kinase-dependent pathways.” Clinical Immunology124(2):170-181.

Glucan containing particles such as yeast cell wall particles can alsobe obtained commercially. Several non-limiting examples include:Nutricell MOS 55 from Biorigin (Sao Paolo, Brazil), SAF-Mannan (SAFAgri, Minneapolis, Minn.), Nutrex (Sensient Technologies, Milwaukee,Wis.), alkali-extracted particles such as those produced by Nutricepts(Nutricepts Inc., Burnsville, Minn.) and ASA Biotech, acid-extracted WGPparticles from Biopolymer Engineering, and organic solvent-extractedparticles such as Adjuvax™ from Alpha-beta Technology, Inc. (Worcester,Mass.) and microparticulate glucan from Novogen (Stamford, Conn.).

Glucan particles such as yeast cell wall particles can have varyinglevels of purity depending on the method of production and/orextraction. In some instances, particles are alkali-extracted,acid-extracted or organic solvent-extracted to remove intracellularcomponents and/or the outer mannoprotein layer of the cell wall. Suchprotocols can produce particles that have a glucan (w/w) content in therange of 50%-90%. In some instances, a particle of lower purity, meaninglower glucan w/w content may be preferred, while in other embodiments, aparticle of higher purity, meaning higher glucan w/w content may bepreferred.

Glucan particles, such as yeast cell wall particles, can have a naturallipid content. For example, the particles can contain 1%, 2%, 3%, 4%,5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%,20% or more than 20% w/w lipid. In the Examples section, theeffectiveness of two glucan particle batches are tested: YGP SAF and YGPSAF+L (containing natural lipids). In some instances, the presence ofnatural lipids may assist in complexation or capture of RNA molecules.

Glucan containing particles typically have a diameter of approximately2-4 microns, although particles with a diameter of less than 2 micronsor greater than 4 microns are also compatible with aspects of theinvention.

The RNA molecule(s) to be delivered are complexed or “trapped” withinthe shell of the glucan particle. The shell or RNA component of theparticle can be labeled for visualization, as described in, andincorporated by reference from, Soto and Ostroff (2008) Bioconjug Chem19:840. Methods of loading GeRPs are discussed further below.

The optimal protocol for uptake of oligonucleotides will depend upon anumber of factors, the most crucial being the type of cells that arebeing used. Other factors that are important in uptake include, but arenot limited to, the nature and concentration of the oligonucleotide, theconfluence of the cells, the type of culture the cells are in (e.g., asuspension culture or plated) and the type of media in which the cellsare grown.

Encapsulating Agents

Encapsulating agents entrap oligonucleotides within vesicles. In anotherembodiment of the invention, an oligonucleotide may be associated with acarrier or vehicle, e.g., liposomes or micelles, although other carrierscould be used, as would be appreciated by one skilled in the art.Liposomes are vesicles made of a lipid bilayer having a structuresimilar to biological membranes. Such carriers are used to facilitatethe cellular uptake or targeting of the oligonucleotide, or improve theoligonucleotides pharmacokinetic or toxicological properties.

For example, the oligonucleotides of the present invention may also beadministered encapsulated in liposomes, pharmaceutical compositionswherein the active ingredient is contained either dispersed or variouslypresent in corpuscles consisting of aqueous concentric layers adherentto lipidic layers. The oligonucleotides, depending upon solubility, maybe present both in the aqueous layer and in the lipidic layer, or inwhat is generally termed a liposomic suspension. The hydrophobic layer,generally but not exclusively, comprises phopholipids such as lecithinand sphingomyelin, steroids such as cholesterol, more or less ionicsurfactants such as diacetylphosphate, stearylamine, or phosphatidicacid, or other materials of a hydrophobic nature. The diameters of theliposomes generally range from about 15 nm to about 5 microns.

The use of liposomes as drug delivery vehicles offers severaladvantages. Liposomes increase intracellular stability, increase uptakeefficiency and improve biological activity. Liposomes are hollowspherical vesicles composed of lipids arranged in a similar fashion asthose lipids which make up the cell membrane. They have an internalaqueous space for entrapping water soluble compounds and range in sizefrom 0.05 to several microns in diameter. Several studies have shownthat liposomes can deliver nucleic acids to cells and that the nucleicacids remain biologically active. For example, a lipid delivery vehicleoriginally designed as a research tool, such as Lipofectin orLIPOFECTAMINE™ 2000, can deliver intact nucleic acid molecules to cells.

Specific advantages of using liposomes include the following: they arenon-toxic and biodegradable in composition; they display longcirculation half-lives; and recognition molecules can be readilyattached to their surface for targeting to tissues. Finally,cost-effective manufacture of liposome-based pharmaceuticals, either ina liquid suspension or lyophilized product, has demonstrated theviability of this technology as an acceptable drug delivery system.

In some aspects, formulations associated with the invention might beselected for a class of naturally occurring or chemically synthesized ormodified saturated and unsaturated fatty acid residues. Fatty acidsmight exist in a form of triglycerides, diglycerides or individual fattyacids. In another embodiment, the use of well-validated mixtures offatty acids and/or fat emulsions currently used in pharmacology forparenteral nutrition may be utilized.

Liposome based formulations are widely used for oligonucleotidedelivery. However, most of commercially available lipid or liposomeformulations contain at least one positively charged lipid (cationiclipids). The presence of this positively charged lipid is believed to beessential for obtaining a high degree of oligonucleotide loading and forenhancing liposome fusogenic properties. Several methods have beenperformed and published to identify optimal positively charged lipidchemistries. However, the commercially available liposome formulationscontaining cationic lipids are characterized by a high level oftoxicity. In vivo limited therapeutic indexes have revealed thatliposome formulations containing positive charged lipids are associatedwith toxicity (i.e. elevation in liver enzymes) at concentrations onlyslightly higher than concentration required to achieve RNA silencing.

Nucleic acids associated with the invention can be hydrophobicallymodified and can be encompassed within neutral nanotransporters. Furtherdescription of neutral nanotransporters is incorporated by referencefrom PCT Application PCT/US2009/005251, filed on Sep. 22, 2009, andentitled “Neutral Nanotransporters.” Such particles enable quantitativeoligonucleotide incorporation into non-charged lipid mixtures. The lackof toxic levels of cationic lipids in such neutral nanotransportercompositions is an important feature.

As demonstrated in PCT/US2009/005251, oligonucleotides can effectivelybe incorporated into a lipid mixture that is free of cationic lipids andsuch a composition can effectively deliver a therapeutic oligonucleotideto a cell in a manner that it is functional. For example, a high levelof activity was observed when the fatty mixture was composed of aphosphatidylcholine base fatty acid and a sterol such as a cholesterol.For instance, one preferred formulation of neutral fatty mixture iscomposed of at least 20% of DOPC or DSPC and at least 20% of sterol suchas cholesterol. Even as low as 1:5 lipid to oligonucleotide ratio wasshown to be sufficient to get complete encapsulation of theoligonucleotide in a non charged formulation.

The neutral nanotransporters compositions enable efficient loading ofoligonucleotide into neutral fat formulation. The composition includesan oligonucleotide that is modified in a manner such that thehydrophobicity of the molecule is increased (for example a hydrophobicmolecule is attached (covalently or no-covalently) to a hydrophobicmolecule on the oligonucleotide terminus or a non-terminal nucleotide,base, sugar, or backbone), the modified oligonucleotide being mixed witha neutral fat formulation (for example containing at least 25% ofcholesterol and 25% of DOPC or analogs thereof). A cargo molecule, suchas another lipid can also be included in the composition. Thiscomposition, where part of the formulation is build into theoligonucleotide itself, enables efficient encapsulation ofoligonucleotide in neutral lipid particles.

In some aspects, stable particles ranging in size from 50 to 140 nm canbe formed upon complexing of hydrophobic oligonucleotides with preferredformulations. It is interesting to mention that the formulation byitself typically does not form small particles, but rather, formsagglomerates, which are transformed into stable 50-120 nm particles uponaddition of the hydrophobic modified oligonucleotide.

The neutral nanotransporter compositions of the invention include ahydrophobic modified polynucleotide, a neutral fatty mixture, andoptionally a cargo molecule. A “hydrophobic modified polynucleotide” asused herein is a polynucleotide of the invention (i.e. sd-rxRNA) thathas at least one modification that renders the polynucleotide morehydrophobic than the polynucleotide was prior to modification. Themodification may be achieved by attaching (covalently or non-covalently)a hydrophobic molecule to the polynucleotide. In some instances thehydrophobic molecule is or includes a lipophilic group.

The term “lipophilic group” means a group that has a higher affinity forlipids than its affinity for water. Examples of lipophilic groupsinclude, but are not limited to, cholesterol, a cholesteryl or modifiedcholesteryl residue, adamantine, dihydrotesterone, long chain alkyl,long chain alkenyl, long chain alkynyl, olely-lithocholic, cholenic,oleoyl-cholenic, palmityl, heptadecyl, myrisityl, bile acids, cholicacid or taurocholic acid, deoxycholate, oleyl litocholic acid, oleoylcholenic acid, glycolipids, phospholipids, sphingolipids, isoprenoids,such as steroids, vitamins, such as vitamin E, fatty acids eithersaturated or unsaturated, fatty acid esters, such as triglycerides,pyrenes, porphyrines, Texaphyrine, adamantane, acridines, biotin,coumarin, fluorescein, rhodamine, Texas-Red, digoxygenin,dimethoxytrityl, t-butyldimethylsilyl, t-butyldiphenylsilyl, cyaninedyes (e.g. Cy3 or Cy5), Hoechst 33258 dye, psoralen, or ibuprofen. Thecholesterol moiety may be reduced (e.g. as in cholestan) or may besubstituted (e.g. by halogen). A combination of different lipophilicgroups in one molecule is also possible.

The hydrophobic molecule may be attached at various positions of thepolynucleotide. As described above, the hydrophobic molecule may belinked to the terminal residue of the polynucleotide such as the 3′ of5′-end of the polynucleotide. Alternatively, it may be linked to aninternal nucleotide or a nucleotide on a branch of the polynucleotide.The hydrophobic molecule may be attached, for instance to a 2′-positionof the nucleotide. The hydrophobic molecule may also be linked to theheterocyclic base, the sugar or the backbone of a nucleotide of thepolynucleotide.

The hydrophobic molecule may be connected to the polynucleotide by alinker moiety. Optionally the linker moiety is a non-nucleotidic linkermoiety. Non-nucleotidic linkers are e.g. abasic residues (dSpacer),oligoethyleneglycol, such as triethyleneglycol (spacer 9) orhexaethylenegylcol (spacer 18), or alkane-diol, such as butanediol. Thespacer units are preferably linked by phosphodiester or phosphorothioatebonds. The linker units may appear just once in the molecule or may beincorporated several times, e.g. via phosphodiester, phosphorothioate,methylphosphonate, or amide linkages.

Typical conjugation protocols involve the synthesis of polynucleotidesbearing an aminolinker at one or more positions of the sequence,however, a linker is not required. The amino group is then reacted withthe molecule being conjugated using appropriate coupling or activatingreagents. The conjugation reaction may be performed either with thepolynucleotide still bound to a solid support or following cleavage ofthe polynucleotide in solution phase. Purification of the modifiedpolynucleotide by HPLC typically results in a pure material.

In some embodiments the hydrophobic molecule is a sterol type conjugate,a PhytoSterol conjugate, cholesterol conjugate, sterol type conjugatewith altered side chain length, fatty acid conjugate, any otherhydrophobic group conjugate, and/or hydrophobic modifications of theinternal nucleoside, which provide sufficient hydrophobicity to beincorporated into micelles.

For purposes of the present invention, the term “sterols”, refers orsteroid alcohols are a subgroup of steroids with a hydroxyl group at the3-position of the A-ring. They are amphipathic lipids synthesized fromacetyl-coenzyme A via the HMG-CoA reductase pathway. The overallmolecule is quite flat. The hydroxyl group on the A ring is polar. Therest of the aliphatic chain is non-polar. Usually sterols are consideredto have an 8 carbon chain at position 17.

For purposes of the present invention, the term “sterol type molecules”,refers to steroid alcohols, which are similar in structure to sterols.The main difference is the structure of the ring and number of carbonsin a position 21 attached side chain.

For purposes of the present invention, the term “PhytoSterols” (alsocalled plant sterols) are a group of steroid alcohols, phytochemicalsnaturally occurring in plants. There are more then 200 different knownPhytoSterols

For purposes of the present invention, the term “Sterol side chain”refers to a chemical composition of a side chain attached at theposition 17 of sterol-type molecule. In a standard definition sterolsare limited to a 4 ring structure carrying a 8 carbon chain at position17. In this invention, the sterol type molecules with side chain longerand shorter than conventional are described. The side chain may branchedor contain double back bones.

Thus, sterols useful in the invention, for example, includecholesterols, as well as unique sterols in which position 17 hasattached side chain of 2-7 or longer then 9 carbons. In a particularembodiment, the length of the polycarbon tail is varied between 5 and 9carbons. Such conjugates may have significantly better in vivo efficacy,in particular delivery to liver. These types of molecules are expectedto work at concentrations 5 to 9 fold lower then oligonucleotidesconjugated to conventional cholesterols.

Alternatively the polynucleotide may be bound to a protein, peptide orpositively charged chemical that functions as the hydrophobic molecule.The proteins may be selected from the group consisting of protamine,dsRNA binding domain, and arginine rich peptides. Exemplary positivelycharged chemicals include spermine, spermidine, cadaverine, andputrescine.

In another embodiment hydrophobic molecule conjugates may demonstrateeven higher efficacy when it is combined with optimal chemicalmodification patterns of the polynucleotide (as described herein indetail), containing but not limited to hydrophobic modifications,phosphorothioate modifications, and 2′ ribo modifications.

In another embodiment the sterol type molecule may be a naturallyoccurring PhytoSterols. The polycarbon chain may be longer than 9 andmay be linear, branched and/or contain double bonds. Some PhytoSterolcontaining polynucleotide conjugates may be significantly more potentand active in delivery of polynucleotides to various tissues. SomePhytoSterols may demonstrate tissue preference and thus be used as a wayto delivery RNAi specifically to particular tissues.

The hydrophobic modified polynucleotide is mixed with a neutral fattymixture to form a micelle. The neutral fatty acid mixture is a mixtureof fats that has a net neutral or slightly net negative charge at oraround physiological pH that can form a micelle with the hydrophobicmodified polynucleotide. For purposes of the present invention, the term“micelle” refers to a small nanoparticle formed by a mixture of noncharged fatty acids and phospholipids. The neutral fatty mixture mayinclude cationic lipids as long as they are present in an amount thatdoes not cause toxicity. In preferred embodiments the neutral fattymixture is free of cationic lipids. A mixture that is free of cationiclipids is one that has less than 1% and preferably 0% of the total lipidbeing cationic lipid. The term “cationic lipid” includes lipids andsynthetic lipids having a net positive charge at or around physiologicalpH. The term “anionic lipid” includes lipids and synthetic lipids havinga net negative charge at or around physiological pH.

The neutral fats bind to the oligonucleotides of the invention by astrong but non-covalent attraction (e.g., an electrostatic, van derWaals, pi-stacking, etc. interaction).

The neutral fat mixture may include formulations selected from a classof naturally occurring or chemically synthesized or modified saturatedand unsaturated fatty acid residues. Fatty acids might exist in a formof triglycerides, diglycerides or individual fatty acids. In anotherembodiment the use of well-validated mixtures of fatty acids and/or fatemulsions currently used in pharmacology for parenteral nutrition may beutilized.

The neutral fatty mixture is preferably a mixture of a choline basedfatty acid and a sterol. Choline based fatty acids include for instance,synthetic phosphocholine derivatives such as DDPC, DLPC, DMPC, DPPC,DSPC, DOPC, POPC, and DEPC. DOPC (chemical registry number 4235-95-4) isdioleoylphosphatidylcholine (also known asdielaidoylphosphatidylcholine, dioleoyl-PC, dioleoylphosphocholine,dioleoyl-sn-glycero-3-phosphocholine, dioleylphosphatidylcholine). DSPC(chemical registry number 816-94-4) is distearoylphosphatidylcholine(also known as 1,2-Distearoyl-sn-Glycero-3-phosphocholine).

The sterol in the neutral fatty mixture may be for instance cholesterol.The neutral fatty mixture may be made up completely of a choline basedfatty acid and a sterol or it may optionally include a cargo molecule.For instance, the neutral fatty mixture may have at least 20% or 25%fatty acid and 20% or 25% sterol.

For purposes of the present invention, the term “Fatty acids” relates toconventional description of fatty acid. They may exist as individualentities or in a form of two- and triglycerides. For purposes of thepresent invention, the term “fat emulsions” refers to safe fatformulations given intravenously to subjects who are unable to getenough fat in their diet. It is an emulsion of soy bean oil (or othernaturally occurring oils) and egg phospholipids. Fat emulsions are beingused for formulation of some insoluble anesthetics. In this disclosure,fat emulsions might be part of commercially available preparations likeIntralipid, Liposyn, Nutrilipid, modified commercial preparations, wherethey are enriched with particular fatty acids or fully denovo-formulated combinations of fatty acids and phospholipids.

In one embodiment, the cells to be contacted with an oligonucleotidecomposition of the invention are contacted with a mixture comprising theoligonucleotide and a mixture comprising a lipid, e.g., one of thelipids or lipid compositions described supra for between about 12 hoursto about 24 hours. In another embodiment, the cells to be contacted withan oligonucleotide composition are contacted with a mixture comprisingthe oligonucleotide and a mixture comprising a lipid, e.g., one of thelipids or lipid compositions described supra for between about 1 andabout five days. In one embodiment, the cells are contacted with amixture comprising a lipid and the oligonucleotide for between aboutthree days to as long as about 30 days. In another embodiment, a mixturecomprising a lipid is left in contact with the cells for at least aboutfive to about 20 days. In another embodiment, a mixture comprising alipid is left in contact with the cells for at least about seven toabout 15 days.

50%-60% of the formulation can optionally be any other lipid ormolecule. Such a lipid or molecule is referred to herein as a cargolipid or cargo molecule. Cargo molecules include but are not limited tointralipid, small molecules, fusogenic peptides or lipids or other smallmolecules might be added to alter cellular uptake, endosomal release ortissue distribution properties. The ability to tolerate cargo moleculesis important for modulation of properties of these particles, if suchproperties are desirable. For instance the presence of some tissuespecific metabolites might drastically alter tissue distributionprofiles. For example use of Intralipid type formulation enriched inshorter or longer fatty chains with various degrees of saturationaffects tissue distribution profiles of these type of formulations (andtheir loads).

An example of a cargo lipid useful according to the invention is afusogenic lipid. For instance, the zwitterionic lipid DOPE (chemicalregistry number 4004-5-1, 1,2-Dioleoyl-sn-Glycero-3-phosphoethanolamine)is a preferred cargo lipid.

Intralipid may be comprised of the following composition: 1 000 mLcontain: purified soybean oil 90 g, purified egg phospholipids 12 g,glycerol anhydrous 22 g, water for injection q.s. ad 1 000 mL. pH isadjusted with sodium hydroxide to pH approximately 8. Energy content/L:4.6 MJ (190 kcal). Osmolality (approx.): 300 mOsm/kg water. In anotherembodiment fat emulsion is Liposyn that contains 5% safflower oil, 5%soybean oil, up to 1.2% egg phosphatides added as an emulsifier and 2.5%glycerin in water for injection. It may also contain sodium hydroxidefor pH adjustment. pH 8.0 (6.0-9.0). Liposyn has an osmolarity of 276 mOsmol/liter (actual).

Variation in the identity, amounts and ratios of cargo lipids affectsthe cellular uptake and tissue distribution characteristics of thesecompounds. For example, the length of lipid tails and level ofsaturability will affect differential uptake to liver, lung, fat andcardiomyocytes. Addition of special hydrophobic molecules like vitaminsor different forms of sterols can favor distribution to special tissueswhich are involved in the metabolism of particular compounds. Complexesare formed at different oligonucleotide concentrations, with higherconcentrations favoring more efficient complex formation.

In another embodiment, the fat emulsion is based on a mixture of lipids.Such lipids may include natural compounds, chemically synthesizedcompounds, purified fatty acids or any other lipids. In yet anotherembodiment the composition of fat emulsion is entirely artificial. In aparticular embodiment, the fat emulsion is more then 70% linoleic acid.In yet another particular embodiment the fat emulsion is at least 1% ofcardiolipin. Linoleic acid (LA) is an unsaturated omega-6 fatty acid. Itis a colorless liquid made of a carboxylic acid with an 18-carbon chainand two cis double bonds.

In yet another embodiment of the present invention, the alteration ofthe composition of the fat emulsion is used as a way to alter tissuedistribution of hydrophobicly modified polynucleotides. This methodologyprovides for the specific delivery of the polynucleotides to particulartissues (FIG. 12).

In another embodiment the fat emulsions of the cargo molecule containmore then 70% of Linoleic acid (C18H32O2) and/or cardiolipin are usedfor specifically delivering RNAi to heart muscle.

Fat emulsions, like intralipid have been used before as a deliveryformulation for some non-water soluble drugs (such as Propofol,re-formulated as Diprivan). Unique features of the present inventioninclude (a) the concept of combining modified polynucleotides with thehydrophobic compound(s), so it can be incorporated in the fat micellesand (b) mixing it with the fat emulsions to provide a reversiblecarrier. After injection into a blood stream, micelles usually bind toserum proteins, including albumin, HDL, LDL and other. This binding isreversible and eventually the fat is absorbed by cells. Thepolynucleotide, incorporated as a part of the micelle will then bedelivered closely to the surface of the cells. After that cellularuptake might be happening though variable mechanisms, including but notlimited to sterol type delivery.

Complexing Agents

Complexing agents bind to the oligonucleotides of the invention by astrong but non-covalent attraction (e.g., an electrostatic, van derWaals, pi-stacking, etc. interaction). In one embodiment,oligonucleotides of the invention can be complexed with a complexingagent to increase cellular uptake of oligonucleotides. An example of acomplexing agent includes cationic lipids. Cationic lipids can be usedto deliver oligonucleotides to cells. However, as discussed above,formulations free in cationic lipids are preferred in some embodiments.

The term “cationic lipid” includes lipids and synthetic lipids havingboth polar and non-polar domains and which are capable of beingpositively charged at or around physiological pH and which bind topolyanions, such as nucleic acids, and facilitate the delivery ofnucleic acids into cells. In general cationic lipids include saturatedand unsaturated alkyl and alicyclic ethers and esters of amines, amides,or derivatives thereof. Straight-chain and branched alkyl and alkenylgroups of cationic lipids can contain, e.g., from 1 to about 25 carbonatoms. Preferred straight chain or branched alkyl or alkene groups havesix or more carbon atoms. Alicyclic groups include cholesterol and othersteroid groups. Cationic lipids can be prepared with a variety ofcounterions (anions) including, e.g., Cl⁻, Br⁻, I⁻, F⁻, acetate,trifluoroacetate, sulfate, nitrite, and nitrate.

Examples of cationic lipids include polyethylenimine, polyamidoamine(PAMAM) starburst dendrimers, Lipofectin (a combination of DOTMA andDOPE), Lipofectase, LIPOFECTAMINE™ (e.g., LIPOFECTAMINE™ 2000), DOPE,Cytofectin (Gilead Sciences, Foster City, Calif.), and Eufectins (JBL,San Luis Obispo, Calif.). Exemplary cationic liposomes can be made fromN-[1-(2,3-dioleoloxy)-propyl]-N,N,N-trimethylammonium chloride (DOTMA),N-[1-(2,3-dioleoloxy)-propyl]-N,N,N-trimethylammonium methylsulfate(DOTAP), 3β-[N—(N′,N′-dimethylaminoethane)carbamoyl]cholesterol(DC-Chol),2,3,-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacetate (DOSPA),1,2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide; anddimethyldioctadecylammonium bromide (DDAB). The cationic lipidN-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA),for example, was found to increase 1000-fold the antisense effect of aphosphorothioate oligonucleotide. (Vlassov et al., 1994, Biochimica etBiophysica Acta 1197:95-108). Oligonucleotides can also be complexedwith, e.g., poly (L-lysine) or avidin and lipids may, or may not, beincluded in this mixture, e.g., steryl-poly (L-lysine).

Cationic lipids have been used in the art to deliver oligonucleotides tocells (see, e.g., U.S. Pat. Nos. 5,855,910; 5,851,548; 5,830,430;5,780,053; 5,767,099; Lewis et al. 1996. Proc. Natl. Acad. Sci. USA93:3176; Hope et al. 1998. Molecular Membrane Biology 15:1). Other lipidcompositions which can be used to facilitate uptake of the instantoligonucleotides can be used in connection with the claimed methods. Inaddition to those listed supra, other lipid compositions are also knownin the art and include, e.g., those taught in U.S. Pat. No. 4,235,871;U.S. Pat. Nos. 4,501,728; 4,837,028; 4,737,323.

In one embodiment lipid compositions can further comprise agents, e.g.,viral proteins to enhance lipid-mediated transfections ofoligonucleotides (Kamata, et al., 1994. Nucl. Acids. Res. 22:536). Inanother embodiment, oligonucleotides are contacted with cells as part ofa composition comprising an oligonucleotide, a peptide, and a lipid astaught, e.g., in U.S. Pat. No. 5,736,392. Improved lipids have also beendescribed which are serum resistant (Lewis, et al., 1996. Proc. Natl.Acad. Sci. 93:3176). Cationic lipids and other complexing agents act toincrease the number of oligonucleotides carried into the cell throughendocytosis.

In another embodiment N-substituted glycine oligonucleotides (peptoids)can be used to optimize uptake of oligonucleotides. Peptoids have beenused to create cationic lipid-like compounds for transfection (Murphy,et al., 1998. Proc. Natl. Acad. Sci. 95:1517). Peptoids can besynthesized using standard methods (e.g., Zuckermann, R. N., et al.1992. J. Am. Chem. Soc. 114:10646; Zuckermann, R. N., et al. 1992. Int.J. Peptide Protein Res. 40:497). Combinations of cationic lipids andpeptoids, liptoids, can also be used to optimize uptake of the subjectoligonucleotides (Hunag, et al., 1998. Chemistry and Biology. 5:345).Liptoids can be synthesized by elaborating peptoid oligonucleotides andcoupling the amino terminal submonomer to a lipid via its amino group(Hunag, et al., 1998. Chemistry and Biology. 5:345).

It is known in the art that positively charged amino acids can be usedfor creating highly active cationic lipids (Lewis et al. 1996. Proc.Natl. Acad. Sci. U.S.A. 93:3176). In one embodiment, a composition fordelivering oligonucleotides of the invention comprises a number ofarginine, lysine, histidine or ornithine residues linked to a lipophilicmoiety (see e.g., U.S. Pat. No. 5,777,153).

In another embodiment, a composition for delivering oligonucleotides ofthe invention comprises a peptide having from between about one to aboutfour basic residues. These basic residues can be located, e.g., on theamino terminal, C-terminal, or internal region of the peptide. Familiesof amino acid residues having similar side chains have been defined inthe art. These families include amino acids with basic side chains(e.g., lysine, arginine, histidine), acidic side chains (e.g., asparticacid, glutamic acid), uncharged polar side chains (e.g., glycine (canalso be considered non-polar), asparagine, glutamine, serine, threonine,tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine,leucine, isoleucine, proline, phenylalanine, methionine, tryptophan),beta-branched side chains (e.g., threonine, valine, isoleucine) andaromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine). Apart from the basic amino acids, a majority or all of theother residues of the peptide can be selected from the non-basic aminoacids, e.g., amino acids other than lysine, arginine, or histidine.Preferably a preponderance of neutral amino acids with long neutral sidechains are used.

In one embodiment, a composition for delivering oligonucleotides of theinvention comprises a natural or synthetic polypeptide having one ormore gamma carboxyglutamic acid residues, or γ-Gla residues. These gammacarboxyglutamic acid residues may enable the polypeptide to bind to eachother and to membrane surfaces. In other words, a polypeptide having aseries of γ-Gla may be used as a general delivery modality that helps anRNAi construct to stick to whatever membrane to which it comes incontact. This may at least slow RNAi constructs from being cleared fromthe blood stream and enhance their chance of homing to the target.

The gamma carboxyglutamic acid residues may exist in natural proteins(for example, prothrombin has 10 γ-Gla residues). Alternatively, theycan be introduced into the purified, recombinantly produced, orchemically synthesized polypeptides by carboxylation using, for example,a vitamin K-dependent carboxylase. The gamma carboxyglutamic acidresidues may be consecutive or non-consecutive, and the total number andlocation of such gamma carboxyglutamic acid residues in the polypeptidecan be regulated/fine tuned to achieve different levels of “stickiness”of the polypeptide.

In one embodiment, the cells to be contacted with an oligonucleotidecomposition of the invention are contacted with a mixture comprising theoligonucleotide and a mixture comprising a lipid, e.g., one of thelipids or lipid compositions described supra for between about 12 hoursto about 24 hours. In another embodiment, the cells to be contacted withan oligonucleotide composition are contacted with a mixture comprisingthe oligonucleotide and a mixture comprising a lipid, e.g., one of thelipids or lipid compositions described supra for between about 1 andabout five days. In one embodiment, the cells are contacted with amixture comprising a lipid and the oligonucleotide for between aboutthree days to as long as about 30 days. In another embodiment, a mixturecomprising a lipid is left in contact with the cells for at least aboutfive to about 20 days. In another embodiment, a mixture comprising alipid is left in contact with the cells for at least about seven toabout 15 days.

For example, in one embodiment, an oligonucleotide composition can becontacted with cells in the presence of a lipid such as cytofectin CS orGSV (available from Glen Research; Sterling, Va.), GS3815, GS2888 forprolonged incubation periods as described herein.

In one embodiment, the incubation of the cells with the mixturecomprising a lipid and an oligonucleotide composition does not reducethe viability of the cells. Preferably, after the transfection periodthe cells are substantially viable. In one embodiment, aftertransfection, the cells are between at least about 70% and at leastabout 100% viable. In another embodiment, the cells are between at leastabout 80% and at least about 95% viable. In yet another embodiment, thecells are between at least about 85% and at least about 90% viable.

In one embodiment, oligonucleotides are modified by attaching a peptidesequence that transports the oligonucleotide into a cell, referred toherein as a “transporting peptide.” In one embodiment, the compositionincludes an oligonucleotide which is complementary to a target nucleicacid molecule encoding the protein, and a covalently attachedtransporting peptide.

The language “transporting peptide” includes an amino acid sequence thatfacilitates the transport of an oligonucleotide into a cell. Exemplarypeptides which facilitate the transport of the moieties to which theyare linked into cells are known in the art, and include, e.g., HIV TATtranscription factor, lactoferrin, Herpes VP22 protein, and fibroblastgrowth factor 2 (Pooga et al. 1998. Nature Biotechnology. 16:857; andDerossi et al. 1998. Trends in Cell Biology. 8:84; Elliott and O'Hare.1997. Cell 88:223).

Oligonucleotides can be attached to the transporting peptide using knowntechniques, e.g., (Prochiantz, A. 1996. Curr. Opin. Neurobiol. 6:629;Derossi et al. 1998. Trends Cell Biol. 8:84; Troy et al. 1996. J.Neurosci. 16:253), Vives et al. 1997. J. Biol. Chem. 272:16010). Forexample, in one embodiment, oligonucleotides bearing an activated thiolgroup are linked via that thiol group to a cysteine present in atransport peptide (e.g., to the cysteine present in the β turn betweenthe second and the third helix of the antennapedia homeodomain astaught, e.g., in Derossi et al. 1998. Trends Cell Biol. 8:84;Prochiantz. 1996. Current Opinion in Neurobiol. 6:629; Allinquant et al.1995. J Cell Biol. 128:919). In another embodiment, a Boc-Cys-(Npys)OHgroup can be coupled to the transport peptide as the last (N-terminal)amino acid and an oligonucleotide bearing an SH group can be coupled tothe peptide (Troy et al. 1996. J. Neurosci. 16:253).

In one embodiment, a linking group can be attached to a nucleomonomerand the transporting peptide can be covalently attached to the linker.In one embodiment, a linker can function as both an attachment site fora transporting peptide and can provide stability against nucleases.Examples of suitable linkers include substituted or unsubstituted C₁-C₂₀alkyl chains, C₂-C₂₀ alkenyl chains, C₂-C₂₀ alkynyl chains, peptides,and heteroatoms (e.g., S, O, NH, etc.). Other exemplary linkers includebifunctional crosslinking agents such assulfosuccinimidyl-4-(maleimidophenyl)-butyrate (SMPB) (see, e.g., Smithet al. Biochem J 1991.276: 417-2).

In one embodiment, oligonucleotides of the invention are synthesized asmolecular conjugates which utilize receptor-mediated endocytoticmechanisms for delivering genes into cells (see, e.g., Bunnell et al.1992. Somatic Cell and Molecular Genetics. 18:559, and the referencescited therein).

Targeting Agents

The delivery of oligonucleotides can also be improved by targeting theoligonucleotides to a cellular receptor. The targeting moieties can beconjugated to the oligonucleotides or attached to a carrier group (i.e.,poly(L-lysine) or liposomes) linked to the oligonucleotides. This methodis well suited to cells that display specific receptor-mediatedendocytosis.

For instance, oligonucleotide conjugates to 6-phosphomannosylatedproteins are internalized 20-fold more efficiently by cells expressingmannose 6-phosphate specific receptors than free oligonucleotides. Theoligonucleotides may also be coupled to a ligand for a cellular receptorusing a biodegradable linker. In another example, the delivery constructis mannosylated streptavidin which forms a tight complex withbiotinylated oligonucleotides. Mannosylated streptavidin was found toincrease 20-fold the internalization of biotinylated oligonucleotides.(Vlassov et al. 1994. Biochimica et Biophysica Acta 1197:95-108).

In addition specific ligands can be conjugated to the polylysinecomponent of polylysine-based delivery systems. For example,transferrin-polylysine, adenovirus-polylysine, and influenza virushemagglutinin HA-2 N-terminal fusogenic peptides-polylysine conjugatesgreatly enhance receptor-mediated DNA delivery in eucaryotic cells.Mannosylated glycoprotein conjugated to poly(L-lysine) in aveolarmacrophages has been employed to enhance the cellular uptake ofoligonucleotides. Liang et al. 1999. Pharmazie 54:559-566.

Because malignant cells have an increased need for essential nutrientssuch as folic acid and transferrin, these nutrients can be used totarget oligonucleotides to cancerous cells. For example, when folic acidis linked to poly(L-lysine) enhanced oligonucleotide uptake is seen inpromyelocytic leukaemia (HL-60) cells and human melanoma (M-14) cells.Ginobbi et al. 1997. Anticancer Res. 17:29. In another example,liposomes coated with maleylated bovine serum albumin, folic acid, orferric protoporphyrin IX, show enhanced cellular uptake ofoligonucleotides in murine macrophages, KB cells, and 2.2.15 humanhepatoma cells. Liang et al. 1999. Pharmazie 54:559-566.

Liposomes naturally accumulate in the liver, spleen, andreticuloendothelial system (so-called, passive targeting). By couplingliposomes to various ligands such as antibodies are protein A, they canbe actively targeted to specific cell populations. For example, proteinA-bearing liposomes may be pretreated with H-2K specific antibodieswhich are targeted to the mouse major histocompatibility complex-encodedH-2K protein expressed on L cells. (Vlassov et al. 1994. Biochimica etBiophysica Acta 1197:95-108).

Other in vitro and/or in vivo delivery of RNAi reagents are known in theart, and can be used to deliver the subject RNAi constructs. See, forexample, U.S. patent application publications 20080152661, 20080112916,20080107694, 20080038296, 20070231392, 20060240093, 20060178327,20060008910, 20050265957, 20050064595, 20050042227, 20050037496,20050026286, 20040162235, 20040072785, 20040063654, 20030157030, WO2008/036825, WO04/065601, and AU2004206255B2, just to name a few (allincorporated by reference).

Administration

The optimal course of administration or delivery of the oligonucleotidesmay vary depending upon the desired result and/or on the subject to betreated. As used herein “administration” refers to contacting cells witholigonucleotides and can be performed in vitro or in vivo. The dosage ofoligonucleotides may be adjusted to optimally reduce expression of aprotein translated from a target nucleic acid molecule, e.g., asmeasured by a readout of RNA stability or by a therapeutic response,without undue experimentation.

For example, expression of the protein encoded by the nucleic acidtarget can be measured to determine whether or not the dosage regimenneeds to be adjusted accordingly. In addition, an increase or decreasein RNA or protein levels in a cell or produced by a cell can be measuredusing any art recognized technique. By determining whether transcriptionhas been decreased, the effectiveness of the oligonucleotide in inducingthe cleavage of a target RNA can be determined.

Any of the above-described oligonucleotide compositions can be usedalone or in conjunction with a pharmaceutically acceptable carrier. Asused herein, “pharmaceutically acceptable carrier” includes appropriatesolvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like. The useof such media and agents for pharmaceutical active substances is wellknown in the art. Except insofar as any conventional media or agent isincompatible with the active ingredient, it can be used in thetherapeutic compositions. Supplementary active ingredients can also beincorporated into the compositions.

Oligonucleotides may be incorporated into liposomes or liposomesmodified with polyethylene glycol or admixed with cationic lipids forparenteral administration. Incorporation of additional substances intothe liposome, for example, antibodies reactive against membrane proteinsfound on specific target cells, can help target the oligonucleotides tospecific cell types.

With respect to in vivo applications, the formulations of the presentinvention can be administered to a patient in a variety of forms adaptedto the chosen route of administration, e.g., parenterally, orally, orintraperitoneally. Parenteral administration, which is preferred,includes administration by the following routes: intravenous;intramuscular; interstitially; intraarterially; subcutaneous; intraocular; intrasynovial; trans epithelial, including transdermal;pulmonary via inhalation; ophthalmic; sublingual and buccal; topically,including ophthalmic; dermal; ocular; rectal; and nasal inhalation viainsufflation. In preferred embodiments, the sd-rxRNA molecules areadministered by intradermal injection or subcutaneously.

Pharmaceutical preparations for parenteral administration includeaqueous solutions of the active compounds in water-soluble orwater-dispersible form. In addition, suspensions of the active compoundsas appropriate oily injection suspensions may be administered. Suitablelipophilic solvents or vehicles include fatty oils, for example, sesameoil, or synthetic fatty acid esters, for example, ethyl oleate ortriglycerides. Aqueous injection suspensions may contain substanceswhich increase the viscosity of the suspension include, for example,sodium carboxymethyl cellulose, sorbitol, or dextran, optionally, thesuspension may also contain stabilizers. The oligonucleotides of theinvention can be formulated in liquid solutions, preferably inphysiologically compatible buffers such as Hank's solution or Ringer'ssolution. In addition, the oligonucleotides may be formulated in solidform and redissolved or suspended immediately prior to use. Lyophilizedforms are also included in the invention.

Pharmaceutical preparations for topical administration includetransdermal patches, ointments, lotions, creams, gels, drops, sprays,suppositories, liquids and powders. In addition, conventionalpharmaceutical carriers, aqueous, powder or oily bases, or thickenersmay be used in pharmaceutical preparations for topical administration.

Pharmaceutical preparations for oral administration include powders orgranules, suspensions or solutions in water or non-aqueous media,capsules, sachets or tablets. In addition, thickeners, flavoring agents,diluents, emulsifiers, dispersing aids, or binders may be used inpharmaceutical preparations for oral administration.

For transmucosal or transdermal administration, penetrants appropriateto the barrier to be permeated are used in the formulation. Suchpenetrants are known in the art, and include, for example, fortransmucosal administration bile salts and fusidic acid derivatives, anddetergents. Transmucosal administration may be through nasal sprays orusing suppositories. For oral administration, the oligonucleotides areformulated into conventional oral administration forms such as capsules,tablets, and tonics. For topical administration, the oligonucleotides ofthe invention are formulated into ointments, salves, gels, or creams asknown in the art.

Drug delivery vehicles can be chosen e.g., for in vitro, for systemic,or for topical administration. These vehicles can be designed to serveas a slow release reservoir or to deliver their contents directly to thetarget cell. An advantage of using some direct delivery drug vehicles isthat multiple molecules are delivered per uptake. Such vehicles havebeen shown to increase the circulation half-life of drugs that wouldotherwise be rapidly cleared from the blood stream. Some examples ofsuch specialized drug delivery vehicles which fall into this categoryare liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, andbioadhesive microspheres.

The described oligonucleotides may be administered systemically to asubject. Systemic absorption refers to the entry of drugs into the bloodstream followed by distribution throughout the entire body.Administration routes which lead to systemic absorption include:intravenous, subcutaneous, intraperitoneal, and intranasal. Each ofthese administration routes delivers the oligonucleotide to accessiblediseased cells. Following subcutaneous administration, the therapeuticagent drains into local lymph nodes and proceeds through the lymphaticnetwork into the circulation. The rate of entry into the circulation hasbeen shown to be a function of molecular weight or size. The use of aliposome or other drug carrier localizes the oligonucleotide at thelymph node. The oligonucleotide can be modified to diffuse into thecell, or the liposome can directly participate in the delivery of eitherthe unmodified or modified oligonucleotide into the cell.

The chosen method of delivery will result in entry into cells. In someembodiments, preferred delivery methods include liposomes (10-400 nm),hydrogels, controlled-release polymers, and other pharmaceuticallyapplicable vehicles, and microinjection or electroporation (for ex vivotreatments).

The pharmaceutical preparations of the present invention may be preparedand formulated as emulsions. Emulsions are usually heterogeneous systemsof one liquid dispersed in another in the form of droplets usuallyexceeding 0.1 μm in diameter. The emulsions of the present invention maycontain excipients such as emulsifiers, stabilizers, dyes, fats, oils,waxes, fatty acids, fatty alcohols, fatty esters, humectants,hydrophilic colloids, preservatives, and anti-oxidants may also bepresent in emulsions as needed. These excipients may be present as asolution in either the aqueous phase, oily phase or itself as a separatephase.

Examples of naturally occurring emulsifiers that may be used in emulsionformulations of the present invention include lanolin, beeswax,phosphatides, lecithin and acacia. Finely divided solids have also beenused as good emulsifiers especially in combination with surfactants andin viscous preparations. Examples of finely divided solids that may beused as emulsifiers include polar inorganic solids, such as heavy metalhydroxides, nonswelling clays such as bentonite, attapulgite, hectorite,kaolin, montrnorillonite, colloidal aluminum silicate and colloidalmagnesium aluminum silicate, pigments and nonpolar solids such as carbonor glyceryl tristearate.

Examples of preservatives that may be included in the emulsionformulations include methyl paraben, propyl paraben, quaternary ammoniumsalts, benzalkonium chloride, esters of p-hydroxybenzoic acid, and boricacid. Examples of antioxidants that may be included in the emulsionformulations include free radical scavengers such as tocopherols, alkylgallates, butylated hydroxyanisole, butylated hydroxytoluene, orreducing agents such as ascorbic acid and sodium metabisulfite, andantioxidant synergists such as citric acid, tartaric acid, and lecithin.

In one embodiment, the compositions of oligonucleotides are formulatedas microemulsions. A microemulsion is a system of water, oil andamphiphile which is a single optically isotropic and thermodynamicallystable liquid solution. Typically microemulsions are prepared by firstdispersing an oil in an aqueous surfactant solution and then adding asufficient amount of a 4th component, generally an intermediatechain-length alcohol to form a transparent system.

Surfactants that may be used in the preparation of microemulsionsinclude, but are not limited to, ionic surfactants, non-ionicsurfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fattyacid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate(MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate(PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate(MO750), decaglycerol sequioleate (S0750), decaglycerol decaoleate(DA0750), alone or in combination with cosurfactants. The cosurfactant,usually a short-chain alcohol such as ethanol, 1-propanol, and1-butanol, serves to increase the interfacial fluidity by penetratinginto the surfactant film and consequently creating a disordered filmbecause of the void space generated among surfactant molecules.

Microemulsions may, however, be prepared without the use ofcosurfactants and alcohol-free self-emulsifying microemulsion systemsare known in the art. The aqueous phase may typically be, but is notlimited to, water, an aqueous solution of the drug, glycerol, PEG300,PEG400, polyglycerols, propylene glycols, and derivatives of ethyleneglycol. The oil phase may include, but is not limited to, materials suchas Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain(C₈-C₁₂) mono, di, and tri-glycerides, polyoxyethylated glyceryl fattyacid esters, fatty alcohols, polyglycolized glycerides, saturatedpolyglycolized C₈-C₁₀ glycerides, vegetable oils and silicone oil.

Microemulsions are particularly of interest from the standpoint of drugsolubilization and the enhanced absorption of drugs. Lipid basedmicroemulsions (both oil/water and water/oil) have been proposed toenhance the oral bioavailability of drugs.

Microemulsions offer improved drug solubilization, protection of drugfrom enzymatic hydrolysis, possible enhancement of drug absorption dueto surfactant-induced alterations in membrane fluidity and permeability,ease of preparation, ease of oral administration over solid dosageforms, improved clinical potency, and decreased toxicity (Constantinideset al., Pharmaceutical Research, 1994, 11:1385; Ho et al., J. Pharm.Sci., 1996, 85:138-143). Microemulsions have also been effective in thetransdermal delivery of active components in both cosmetic andpharmaceutical applications. It is expected that the microemulsioncompositions and formulations of the present invention will facilitatethe increased systemic absorption of oligonucleotides from thegastrointestinal tract, as well as improve the local cellular uptake ofoligonucleotides within the gastrointestinal tract, vagina, buccalcavity and other areas of administration.

In an embodiment, the present invention employs various penetrationenhancers to affect the efficient delivery of nucleic acids,particularly oligonucleotides, to the skin of animals. Evennon-lipophilic drugs may cross cell membranes if the membrane to becrossed is treated with a penetration enhancer. In addition toincreasing the diffusion of non-lipophilic drugs across cell membranes,penetration enhancers also act to enhance the permeability of lipophilicdrugs.

Five categories of penetration enhancers that may be used in the presentinvention include: surfactants, fatty acids, bile salts, chelatingagents, and non-chelating non-surfactants. Other agents may be utilizedto enhance the penetration of the administered oligonucleotides include:glycols such as ethylene glycol and propylene glycol, pyrrols such as2-15 pyrrol, azones, and terpenes such as limonene, and menthone.

The oligonucleotides, especially in lipid formulations, can also beadministered by coating a medical device, for example, a catheter, suchas an angioplasty balloon catheter, with a cationic lipid formulation.Coating may be achieved, for example, by dipping the medical device intoa lipid formulation or a mixture of a lipid formulation and a suitablesolvent, for example, an aqueous-based buffer, an aqueous solvent,ethanol, methylene chloride, chloroform and the like. An amount of theformulation will naturally adhere to the surface of the device which issubsequently administered to a patient, as appropriate. Alternatively, alyophilized mixture of a lipid formulation may be specifically bound tothe surface of the device. Such binding techniques are described, forexample, in K. Ishihara et al., Journal of Biomedical MaterialsResearch, Vol. 27, pp. 1309-1314 (1993), the disclosures of which areincorporated herein by reference in their entirety.

The useful dosage to be administered and the particular mode ofadministration will vary depending upon such factors as the cell type,or for in vivo use, the age, weight and the particular animal and regionthereof to be treated, the particular oligonucleotide and deliverymethod used, the therapeutic or diagnostic use contemplated, and theform of the formulation, for example, suspension, emulsion, micelle orliposome, as will be readily apparent to those skilled in the art.Typically, dosage is administered at lower levels and increased untilthe desired effect is achieved. When lipids are used to deliver theoligonucleotides, the amount of lipid compound that is administered canvary and generally depends upon the amount of oligonucleotide agentbeing administered. For example, the weight ratio of lipid compound tooligonucleotide agent is preferably from about 1:1 to about 15:1, with aweight ratio of about 5:1 to about 10:1 being more preferred. Generally,the amount of cationic lipid compound which is administered will varyfrom between about 0.1 milligram (mg) to about 1 gram (g). By way ofgeneral guidance, typically between about 0.1 mg and about 10 mg of theparticular oligonucleotide agent, and about 1 mg to about 100 mg of thelipid compositions, each per kilogram of patient body weight, isadministered, although higher and lower amounts can be used.

The agents of the invention are administered to subjects or contactedwith cells in a biologically compatible form suitable for pharmaceuticaladministration. By “biologically compatible form suitable foradministration” is meant that the oligonucleotide is administered in aform in which any toxic effects are outweighed by the therapeuticeffects of the oligonucleotide. In one embodiment, oligonucleotides canbe administered to subjects. Examples of subjects include mammals, e.g.,humans and other primates; cows, pigs, horses, and farming(agricultural) animals; dogs, cats, and other domesticated pets; mice,rats, and transgenic non-human animals.

Administration of an active amount of an oligonucleotide of the presentinvention is defined as an amount effective, at dosages and for periodsof time necessary to achieve the desired result. For example, an activeamount of an oligonucleotide may vary according to factors such as thetype of cell, the oligonucleotide used, and for in vivo uses the diseasestate, age, sex, and weight of the individual, and the ability of theoligonucleotide to elicit a desired response in the individual.Establishment of therapeutic levels of oligonucleotides within the cellis dependent upon the rates of uptake and efflux or degradation.Decreasing the degree of degradation prolongs the intracellularhalf-life of the oligonucleotide. Thus, chemically-modifiedoligonucleotides, e.g., with modification of the phosphate backbone, mayrequire different dosing.

The exact dosage of an oligonucleotide and number of doses administeredwill depend upon the data generated experimentally and in clinicaltrials. Several factors such as the desired effect, the deliveryvehicle, disease indication, and the route of administration, willaffect the dosage. Dosages can be readily determined by one of ordinaryskill in the art and formulated into the subject pharmaceuticalcompositions. Preferably, the duration of treatment will extend at leastthrough the course of the disease symptoms.

Dosage regimens may be adjusted to provide the optimum therapeuticresponse. For example, the oligonucleotide may be repeatedlyadministered, e.g., several doses may be administered daily or the dosemay be proportionally reduced as indicated by the exigencies of thetherapeutic situation. One of ordinary skill in the art will readily beable to determine appropriate doses and schedules of administration ofthe subject oligonucleotides, whether the oligonucleotides are to beadministered to cells or to subjects.

Administration of sd-rxRNAs, such as through intradermal injection orsubcutaneous delivery, can be optimized through testing of dosingregimens. In some embodiments, a single administration is sufficient. Tofurther prolong the effect of the administered sd-rxRNA, the sd-rxRNAcan be administered in a slow-release formulation or device, as would befamiliar to one of ordinary skill in the art. The hydrophobic nature ofsd-rxRNA compounds can enable use of a wide variety of polymers, some ofwhich are not compatible with conventional oligonucleotide delivery.

In other embodiments, the sd-rxRNA is administered multiple times. Insome instances it is administered daily, bi-weekly, weekly, every twoweeks, every three weeks, monthly, every two months, every three months,every four months, every five months, every six months or lessfrequently than every six months. In some instances, it is administeredmultiple times per day, week, month and/or year. For example, it can beadministered approximately every hour, 2 hours, 3 hours, 4 hours, 5hours, 6 hours, 7 hours, 8 hours, 9 hours 10 hours, 12 hours or morethan twelve hours. It can be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10or more than 10 times per day.

Aspects of the invention relate to administering sd-rxRNA molecules to asubject. In some instances the subject is a patient and administeringthe sd-rxRNA molecule involves administering the sd-rxRNA molecule in adoctor's office.

In some embodiments, more than one sd-rxRNA molecule is administeredsimultaneously. For example a composition may be administered thatcontains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 differentsd-rxRNA molecules. In certain embodiments, a composition comprises 2 or3 different sd-rxRNA molecules. When a composition comprises more thanone sd-rxRNA, the sd-rxRNA molecules within the composition can bedirected to the same gene or to different genes.

FIG. 1 reveals the expression profile for several genes associated withthe invention. As expected, target gene expression is elevated early andreturns to normal by day 10. FIG. 2 provides a summary of experimentaldesign. FIGS. 3-6 show in vivo silencing of MAP4K4 and PPIB expressionfollowing intradermal injection of sd-rxRNA molecules targeting thesegenes. FIGS. 7-8 show that the silencing effect of sd-rxRNAs can persistfor at least 8 days. Thus, in some embodiments, sd-rxRNA is administeredwithin 8 days prior to an event that compromises or damages the skinsuch as a surgery. For examples, an sd-rxRNA could be administered 1, 2,3, 4, 5, 6, 7, 8, 9, 10 or more than 10 days prior to an event thatcompromises or damages the skin. FIG. 9 demonstrates examples of dosingregimens.

In some instances, the effective amount of sd-rxRNA that is delivered bysubcutaneous administration is at least approximately 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,98, 99, 100 or more than 100 mg/kg including any intermediate values.

In some instances, the effective amount of sd-rxRNA that is deliveredthrough intradermal injection is at least approximately 1, 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100,125, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750,800, 850, 900, 950 or more than 950 μg including any intermediatevalues.

sd-rxRNA molecules administered through methods described herein areeffectively targeted to all the cell types in the skin.

Physical methods of introducing nucleic acids include injection of asolution containing the nucleic acid, bombardment by particles coveredby the nucleic acid, soaking the cell or organism in a solution of thenucleic acid, or electroporation of cell membranes in the presence ofthe nucleic acid. A viral construct packaged into a viral particle wouldaccomplish both efficient introduction of an expression construct intothe cell and transcription of nucleic acid encoded by the expressionconstruct. Other methods known in the art for introducing nucleic acidsto cells may be used, such as lipid-mediated carrier transport,chemical-mediated transport, such as calcium phosphate, and the like.Thus the nucleic acid may be introduced along with components thatperform one or more of the following activities: enhance nucleic aciduptake by the cell, inhibit annealing of single strands, stabilize thesingle strands, or other-wise increase inhibition of the target gene.

Nucleic acid may be directly introduced into the cell (i.e.,intracellularly); or introduced extracellularly into a cavity,interstitial space, into the circulation of an organism, introducedorally, or may be introduced by bathing a cell or organism in a solutioncontaining the nucleic acid. Vascular or extravascular circulation, theblood or lymph system, and the cerebrospinal fluid are sites where thenucleic acid may be introduced.

The cell with the target gene may be derived from or contained in anyorganism. The organism may a plant, animal, protozoan, bacterium, virus,or fungus. The plant may be a monocot, dicot or gymnosperm; the animalmay be a vertebrate or invertebrate. Preferred microbes are those usedin agriculture or by industry, and those that are pathogenic for plantsor animals.

Alternatively, vectors, e.g., transgenes encoding a siRNA of theinvention can be engineered into a host cell or transgenic animal usingart recognized techniques.

A further preferred use for the agents of the present invention (orvectors or transgenes encoding same) is a functional analysis to becarried out in eukaryotic cells, or eukaryotic non-human organisms,preferably mammalian cells or organisms and most preferably human cells,e.g. cell lines such as HeLa or 293 or rodents, e.g. rats and mice. Byadministering a suitable priming agent/RNAi agent which is sufficientlycomplementary to a target mRNA sequence to direct target-specific RNAinterference, a specific knockout or knockdown phenotype can be obtainedin a target cell, e.g. in cell culture or in a target organism.

Thus, a further subject matter of the invention is a eukaryotic cell ora eukaryotic non-human organism exhibiting a target gene-specificknockout or knockdown phenotype comprising a fully or at least partiallydeficient expression of at least one endogenous target gene wherein saidcell or organism is transfected with at least one vector comprising DNAencoding an RNAi agent capable of inhibiting the expression of thetarget gene. It should be noted that the present invention allows atarget-specific knockout or knockdown of several different endogenousgenes due to the specificity of the RNAi agent.

Gene-specific knockout or knockdown phenotypes of cells or non-humanorganisms, particularly of human cells or non-human mammals may be usedin analytic to procedures, e.g. in the functional and/or phenotypicalanalysis of complex physiological processes such as analysis of geneexpression profiles and/or proteomes. Preferably the analysis is carriedout by high throughput methods using oligonucleotide based chips.

Assays of Oligonucleotide Stability

In some embodiments, the oligonucleotides of the invention arestabilized, i.e., substantially resistant to endonuclease andexonuclease degradation. An oligonucleotide is defined as beingsubstantially resistant to nucleases when it is at least about 3-foldmore resistant to attack by an endogenous cellular nuclease, and ishighly nuclease resistant when it is at least about 6-fold moreresistant than a corresponding oligonucleotide. This can be demonstratedby showing that the oligonucleotides of the invention are substantiallyresistant to nucleases using techniques which are known in the art.

One way in which substantial stability can be demonstrated is by showingthat the oligonucleotides of the invention function when delivered to acell, e.g., that they reduce transcription or translation of targetnucleic acid molecules, e.g., by measuring protein levels or bymeasuring cleavage of mRNA. Assays which measure the stability of targetRNA can be performed at about 24 hours post-transfection (e.g., usingNorthern blot techniques, RNase Protection Assays, or QC-PCR assays asknown in the art). Alternatively, levels of the target protein can bemeasured. Preferably, in addition to testing the RNA or protein levelsof interest, the RNA or protein levels of a control, non-targeted genewill be measured (e.g., actin, or preferably a control with sequencesimilarity to the target) as a specificity control. RNA or proteinmeasurements can be made using any art-recognized technique. Preferably,measurements will be made beginning at about 16-24 hours posttransfection. (M. Y. Chiang, et al. 1991. J Biol Chem. 266:18162-71; T.Fisher, et al. 1993. Nucleic Acids Research. 21 3857).

The ability of an oligonucleotide composition of the invention toinhibit protein synthesis can be measured using techniques which areknown in the art, for example, by detecting an inhibition in genetranscription or protein synthesis. For example, Nuclease S1 mapping canbe performed. In another example, Northern blot analysis can be used tomeasure the presence of RNA encoding a particular protein. For example,total RNA can be prepared over a cesium chloride cushion (see, e.g.,Ausebel et al., 1987. Current Protocols in Molecular Biology (Greene &Wiley, New York)). Northern blots can then be made using the RNA andprobed (see, e.g., Id.). In another example, the level of the specificmRNA produced by the target protein can be measured, e.g., using PCR. Inyet another example, Western blots can be used to measure the amount oftarget protein present. In still another embodiment, a phenotypeinfluenced by the amount of the protein can be detected. Techniques forperforming Western blots are well known in the art, see, e.g., Chen etal. J. Biol. Chem. 271:28259.

In another example, the promoter sequence of a target gene can be linkedto a reporter gene and reporter gene transcription (e.g., as describedin more detail below) can be monitored. Alternatively, oligonucleotidecompositions that do not target a promoter can be identified by fusing aportion of the target nucleic acid molecule with a reporter gene so thatthe reporter gene is transcribed. By monitoring a change in theexpression of the reporter gene in the presence of the oligonucleotidecomposition, it is possible to determine the effectiveness of theoligonucleotide composition in inhibiting the expression of the reportergene. For example, in one embodiment, an effective oligonucleotidecomposition will reduce the expression of the reporter gene.

A “reporter gene” is a nucleic acid that expresses a detectable geneproduct, which may be RNA or protein. Detection of mRNA expression maybe accomplished by Northern blotting and detection of protein may beaccomplished by staining with antibodies specific to the protein.Preferred reporter genes produce a readily detectable product. Areporter gene may be operably linked with a regulatory DNA sequence suchthat detection of the reporter gene product provides a measure of thetranscriptional activity of the regulatory sequence. In preferredembodiments, the gene product of the reporter gene is detected by anintrinsic activity associated with that product. For instance, thereporter gene may encode a gene product that, by enzymatic activity,gives rise to a detectable signal based on color, fluorescence, orluminescence. Examples of reporter genes include, but are not limitedto, those coding for chloramphenicol acetyl transferase (CAT),luciferase, beta-galactosidase, and alkaline phosphatase.

One skilled in the art would readily recognize numerous reporter genessuitable for use in the present invention. These include, but are notlimited to, chloramphenicol acetyltransferase (CAT), luciferase, humangrowth hormone (hGH), and beta-galactosidase. Examples of such reportergenes can be found in F. A. Ausubel et al., Eds., Current Protocols inMolecular Biology, John Wiley & Sons, New York, (1989). Any gene thatencodes a detectable product, e.g., any product having detectableenzymatic activity or against which a specific antibody can be raised,can be used as a reporter gene in the present methods.

One reporter gene system is the firefly luciferase reporter system.(Gould, S. J., and Subramani, S. 1988. Anal. Biochem., 7:404-408incorporated herein by reference). The luciferase assay is fast andsensitive. In this assay, a lysate of the test cell is prepared andcombined with ATP and the substrate luciferin. The encoded enzymeluciferase catalyzes a rapid, ATP dependent oxidation of the substrateto generate a light-emitting product. The total light output is measuredand is proportional to the amount of luciferase present over a widerange of enzyme concentrations.

CAT is another frequently used reporter gene system; a major advantageof this system is that it has been an extensively validated and iswidely accepted as a measure of promoter activity. (Gorman C. M.,Moffat, L. F., and Howard, B. H. 1982. Mol. Cell. Biol., 2:1044-1051).In this system, test cells are transfected with CAT expression vectorsand incubated with the candidate substance within 2-3 days of theinitial transfection. Thereafter, cell extracts are prepared. Theextracts are incubated with acetyl CoA and radioactive chloramphenicol.Following the incubation, acetylated chloramphenicol is separated fromnonacetylated form by thin layer chromatography. In this assay, thedegree of acetylation reflects the CAT gene activity with the particularpromoter.

Another suitable reporter gene system is based on immunologic detectionof hGH. This system is also quick and easy to use. (Selden, R.,Burke-Howie, K. Rowe, M. E., Goodman, H. M., and Moore, D. D. (1986),Mol. Cell, Biol., 6:3173-3179 incorporated herein by reference). The hGHsystem is advantageous in that the expressed hGH polypeptide is assayedin the media, rather than in a cell extract. Thus, this system does notrequire the destruction of the test cells. It will be appreciated thatthe principle of this reporter gene system is not limited to hGH butrather adapted for use with any polypeptide for which an antibody ofacceptable specificity is available or can be prepared.

In one embodiment, nuclease stability of a double-strandedoligonucleotide of the invention is measured and compared to a control,e.g., an RNAi molecule typically used in the art (e.g., a duplexoligonucleotide of less than 25 nucleotides in length and comprising 2nucleotide base overhangs) or an unmodified RNA duplex with blunt ends.

The target RNA cleavage reaction achieved using the siRNAs of theinvention is highly sequence specific. Sequence identity may determinedby sequence comparison and alignment algorithms known in the art. Todetermine the percent identity of two nucleic acid sequences (or of twoamino acid sequences), the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in the first sequence or secondsequence for optimal alignment). A preferred, non-limiting example of alocal alignment algorithm utilized for the comparison of sequences isthe algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA87:2264-68, modified as in Karlin and Altschul (1993) Proc. Natl. Acad.Sci. USA 90:5873-77. Such an algorithm is incorporated into the BLASTprograms (version 2.0) of Altschul, et al. (1990) J. Mol. Biol.215:403-10. Greater than 90% sequence identity, e.g., 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or even 100% sequence identity, between thesiRNA and the portion of the target gene is preferred. Alternatively,the siRNA may be defined functionally as a nucleotide sequence (oroligonucleotide sequence) that is capable of hybridizing with a portionof the target gene transcript. Examples of stringency conditions forpolynucleotide hybridization are provided in Sambrook, J., E. F.Fritsch, and T. Maniatis, 1989, Molecular Cloning: A Laboratory Manual,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., chapters9 and 11, and Current Protocols in Molecular Biology, 1995, F. M.Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and6.3-6.4, incorporated herein by reference.

Therapeutic Use

By inhibiting the expression of a gene, the oligonucleotide compositionsof the present invention can be used to treat any disease involving theexpression of a protein. Examples of diseases that can be treated byoligonucleotide compositions, just to illustrate, include: cancer,retinopathies, autoimmune diseases, inflammatory diseases (i.e., ICAM-1related disorders, Psoriasis, Ulcerative Colitus, Crohn's disease),viral diseases (i.e., HIV, Hepatitis C), miRNA disorders, andcardiovascular diseases.

In one embodiment, in vitro treatment of cells with oligonucleotides canbe used for ex vivo therapy of cells removed from a subject (e.g., fortreatment of leukemia or viral infection) or for treatment of cellswhich did not originate in the subject, but are to be administered tothe subject (e.g., to eliminate transplantation antigen expression oncells to be transplanted into a subject). In addition, in vitrotreatment of cells can be used in non-therapeutic settings, e.g., toevaluate gene function, to study gene regulation and protein synthesisor to evaluate improvements made to oligonucleotides designed tomodulate gene expression or protein synthesis. In vivo treatment ofcells can be useful in certain clinical settings where it is desirableto inhibit the expression of a protein. There are numerous medicalconditions for which antisense therapy is reported to be suitable (see,e.g., U.S. Pat. No. 5,830,653) as well as respiratory syncytial virusinfection (WO 95/22,553) influenza virus (WO 94/23,028), andmalignancies (WO 94/08,003). Other examples of clinical uses ofantisense sequences are reviewed, e.g., in Glaser. 1996. GeneticEngineering News 16:1. Exemplary targets for cleavage byoligonucleotides include, e.g., protein kinase Ca, ICAM-1, c-raf kinase,p53, c-myb, and the bcr/abl fusion gene found in chronic myelogenousleukemia.

The subject nucleic acids can be used in RNAi-based therapy in anyanimal having RNAi pathway, such as human, non-human primate, non-humanmammal, non-human vertebrates, rodents (mice, rats, hamsters, rabbits,etc.), domestic livestock animals, pets (cats, dogs, etc.), Xenopus,fish, insects (Drosophila, etc.), and worms (C. elegans), etc.

The invention provides methods for preventing in a subject, a disease orcondition associated with an aberrant or unwanted target gene expressionor activity, by administering to the subject a therapeutic agent (e.g.,a RNAi agent or vector or transgene encoding same). If appropriate,subjects are first treated with a priming agent so as to be moreresponsive to the subsequent RNAi therapy. Subjects at risk for adisease which is caused or contributed to by aberrant or unwanted targetgene expression or activity can be identified by, for example, any or acombination of diagnostic or prognostic assays as described herein.Administration of a prophylactic agent can occur prior to themanifestation of symptoms characteristic of the target gene aberrancy,such that a disease or disorder is prevented or, alternatively, delayedin its progression. Depending on the type of target gene aberrancy, forexample, a target gene, target gene agonist or target gene antagonistagent can be used for treating the subject.

In another aspect, the invention pertains to methods of modulatingtarget gene expression, protein expression or activity for therapeuticpurposes. Accordingly, in an exemplary embodiment, the modulatory methodof the invention involves contacting a cell capable of expressing targetgene with a therapeutic agent of the invention that is specific for thetarget gene or protein (e.g., is specific for the mRNA encoded by saidgene or specifying the amino acid sequence of said protein) such thatexpression or one or more of the activities of target protein ismodulated. These modulatory methods can be performed in vitro (e.g., byculturing the cell with the agent), in vivo (e.g., by administering theagent to a subject), or ex vivo. Typically, subjects are first treatedwith a priming agent so as to be more responsive to the subsequent RNAitherapy. As such, the present invention provides methods of treating anindividual afflicted with a disease or disorder characterized byaberrant or unwanted expression or activity of a target gene polypeptideor nucleic acid molecule. Inhibition of target gene activity isdesirable in situations in which target gene is abnormally unregulatedand/or in which decreased target gene activity is likely to have abeneficial effect.

The therapeutic agents of the invention can be administered toindividuals to treat (prophylactically or therapeutically) disordersassociated with aberrant or unwanted target gene activity. Inconjunction with such treatment, pharmacogenomics (i.e., the study ofthe relationship between an individual's genotype and that individual'sresponse to a foreign compound or drug) may be considered. Differencesin metabolism of therapeutics can lead to severe toxicity or therapeuticfailure by altering the relation between dose and blood concentration ofthe pharmacologically active drug. Thus, a physician or clinician mayconsider applying knowledge obtained in relevant pharmacogenomicsstudies in determining whether to administer a therapeutic agent as wellas tailoring the dosage and/or therapeutic regimen of treatment with atherapeutic agent. Pharmacogenomics deals with clinically significanthereditary variations in the response to drugs due to altered drugdisposition and abnormal action in affected persons. See, for example,Eichelbaum, M. et al. (1996) Clin. Exp. Pharmacol. Physiol. 23(10-11):983-985 and Linder, M. W. et al. (1997) Clin. Chem. 43(2):254-266

RNAi in Skin Indications

Nucleic acid molecules, or compositions comprising nucleic acidmolecules, described herein may in some embodiments be administered topre-treat, treat or prevent compromised skin. As used herein“compromised skin” refers to skin which exhibits characteristicsdistinct from normal skin. Compromised skin may occur in associationwith a dermatological condition. Several non-limiting examples ofdermatological conditions include rosacea, common acne, seborrheicdermatitis, perioral dermatitis, acneform rashes, transient acantholyticdermatosis, and acne necrotica miliaris. In some instances, compromisedskin may comprise a wound and/or scar tissue. In some instances, methodsand compositions associated with the invention may be used to promotewound healing, prevention, reduction or inhibition of scarring, and/orpromotion of re-epithelialisation of wounds.

A subject can be pre-treated or treated prophylactically with a moleculeassociated with the invention, prior to the skin of the subject becomingcompromised. As used herein “pre-treatment” or “prophylactic treatment”refers to administering a nucleic acid to the skin prior to the skinbecoming compromised. For example, a subject could be pre-treated 15minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 24hours, 48 hours, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days or morethan 8 days prior to the skin becoming compromised. In otherembodiments, a subject can be treated with a molecule associated withthe invention immediately before the skin becomes compromised and/orsimultaneous to the skin becoming compromised and/or after the skin hasbeen compromised. In some embodiments, the skin is compromised through amedical procedure such as surgery, including elective surgery. Incertain embodiments methods and compositions may be applied to areas ofthe skin that are believed to be at risk of becoming compromised. Itshould be appreciated that one of ordinary skill in the art would beable to optimize timing of administration using no more than routineexperimentation.

In some aspects, methods associated with the invention can be applied topromote healing of compromised skin. Administration can occur at anytime up until the compromised skin has healed, even if the compromisedskin has already partially healed. The timing of administration candepend on several factors including the nature of the compromised skin,the degree of damage within the compromised skin, and the size of thecompromised area. In some embodiments administration may occurimmediately after the skin is compromised, or 30 minutes, 1 hour, 2hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours, 48 hours, or morethan 48 hours after the skin has been compromised. Methods andcompositions of the invention may be administered one or more times asnecessary. For example, in some embodiments, compositions may beadministered daily or twice daily. In some instances, compositions maybe administered both before and after formation of compromised skin.

Compositions associated with the invention may be administered by anysuitable route. In some embodiments, administration occurs locally at anarea of compromised skin. For example, compositions may be administeredby intradermal injection. Compositions for intradermal injection mayinclude injectable solutions. Intradermal injection may in someembodiments occur around the are of compromised skin or at a site wherethe skin is likely to become compromised. In some embodiments,compositions may also be administered in a topical form, such as in acream or ointment. In some embodiments, administration of compositionsdescribed herein comprises part of an initial treatment or pre-treatmentof compromised skin, while in other embodiments, administration of suchcompositions comprises follow-up care for an area of compromised skin.

The appropriate amount of a composition or medicament to be applied candepend on many different factors and can be determined by one ofordinary skill in the art through routine experimentation. Severalnon-limiting factors that might be considered include biologicalactivity and bioavailability of the agent, nature of the agent, mode ofadministration, half-life, and characteristics of the subject to betreated.

In some aspects, nucleic acid molecules associated with the inventionmay also be used in treatment and/or prevention of fibrotic disorders,including pulmonary fibrosis, liver cirrhosis, scleroderma andglomerulonephritis, lung fibrosis, liver fibrosis, skin fibrosis, musclefibrosis, radiation fibrosis, kidney fibrosis, proliferativevitreoretinopathy, restenosis, and uterine fibrosis.

A therapeutically effective amount of a nucleic acid molecule describedherein may in some embodiments be an amount sufficient to prevent theformation of compromised skin and/or improve the condition ofcompromised skin and/or to treat or prevent a fibrotic disorder. In someembodiments, improvement of the condition of compromised skin maycorrespond to promotion of wound healing and/or inhibition of scarringand/or promotion of epithelial regeneration. The extent of prevention offormation of compromised skin and/or improvement to the condition ofcompromised skin may in some instances be determined by, for example, adoctor or clinician.

The ability of nucleic acid molecules associated with the invention toprevent the formation of compromised skin and/or improve the conditionof compromised skin may in some instances be measured with reference toproperties exhibited by the skin. In some instances, these propertiesmay include rate of epithelialisation and/or decreased size of an areaof compromised skin compared to control skin at comparable time points.

As used herein, prevention of formation of compromised skin, for exampleprior to a surgical procedure, and/or improvement of the condition ofcompromised skin, for example after a surgical procedure, can encompassany increase in the rate of healing in the compromised skin as comparedwith the rate of healing occurring in a control sample. In someinstances, the condition of compromised skin may be assessed withrespect to either comparison of the rate of re-epithelialisationachieved in treated and control skin, or comparison of the relativeareas of treated and control areas of compromised skin at comparabletime points. In some aspects, a molecule that prevents formation ofcompromised skin or promotes healing of compromised skin may be amolecule that, upon administration, causes the area of compromised skinto exhibit an increased rate of re-epithelialisation and/or a reductionof the size of compromised skin compared to a control at comparable timepoints. In some embodiments, the healing of compromised skin may giverise to a rate of healing that is 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, 90% or 100% greater than the rate occurring in controls.

In some aspects, subjects to be treated by methods and compositionsassociated with the invention may be subjects who will undergo, areundergoing or have undergone a medical procedure such as a surgery. Insome embodiments, the subject may be prone to defective, delayed orotherwise impaired re-epithelialisation, such as dermal wounds in theaged. Other non-limiting examples of conditions or disorders in whichwound healing is associated with delayed or otherwise impairedre-epithelialisation include patients suffering from diabetes, patientswith polypharmacy, post-menopausal women, patients susceptible topressure injuries, patients with venous disease, clinically obesepatients, patients receiving chemotherapy, patients receivingradiotherapy, patients receiving steroid treatment, andimmuno-compromised patients. In some instances, defectivere-epithelialisation response can contributes to infections at the woundsite, and to the formation of chronic wounds such as ulcers.

In some embodiments, methods associated with the invention may promotethe re-epithelialisation of compromised skin in chronic wounds, such asulcers, and may also inhibit scarring associated with wound healing. Inother embodiments, methods associated with the invention are applied toprevention or treatment of compromised skin in acute wounds in patientspredisposed to impaired wound healing developing into chronic wounds. Inother aspects, methods associated with the invention are applied topromote accelerated healing of compromised skin while preventing,reducing or inhibiting scarring for use in general clinical contexts. Insome aspects, this can involve the treatment of surgical incisions andapplication of such methods may result in the prevention, reduction orinhibition of scarring that may otherwise occur on such healing. Suchtreatment may result in the scars being less noticeable and exhibitingregeneration of a more normal skin structure. In other embodiments, thecompromised skin that is treated is not compromised skin that is causedby a surgical incision. The compromised skin may be subject to continuedcare and continued application of medicaments to encouragere-epithelialisation and healing.

In some aspects, methods associated with the invention may also be usedin the treatment of compromised skin associated with graftingprocedures. This can involve treatment at a graft donor site and/or at agraft recipient site. Grafts can in some embodiments involve skin,artificial skin, or skin substitutes. Methods associated with theinvention can also be used for promoting epithelial regeneration. Asused herein, promotion of epithelial regeneration encompasses anyincrease in the rate of epithelial regeneration as compared to theregeneration occurring in a control-treated or untreated epithelium. Therate of epithelial regeneration attained can in some instances becompared with that taking place in control-treated or untreatedepithelia using any suitable model of epithelial regeneration known inthe art. Promotion of epithelial regeneration may be of use to induceeffective re-epithelialisation in contexts in which there-epithelialisation response is impaired, inhibited, retarded orotherwise defective. Promotion of epithelial regeneration may be alsoeffected to accelerate the rate of defective or normal epithelialregeneration responses in patients suffering from epithelial damage.

Some instances where re-epithelialisation response may be defectiveinclude conditions such as pemphigus, Hailey-Hailey disease (familialbenign pemphigus), toxic epidermal necrolysis (TEN)/Lyell's syndrome,epidermolysis bullosa, cutaneous leishmaniasis and actinic keratosis.Defective re-epithelialisation of the lungs may be associated withidiopathic pulmonary fibrosis (IPF) or interstitial lung disease.Defective re-epithelialisation of the eye may be associated withconditions such as partial limbal stem cell deficiency or cornealerosions. Defective re-epithelialisation of the gastrointestinal tractor colon may be associated with conditions such as chronic anal fissures(fissure in ano), ulcerative colitis or Crohn's disease, and otherinflammatory bowel disorders.

In some aspects, methods associated with the invention are used toprevent, reduce or otherwise inhibit compromised skin associated withscarring. This can be applied to any site within the body and any tissueor organ, including the skin, eye, nerves, tendons, ligaments, muscle,and oral cavity (including the lips and palate), as well as internalorgans (such as the liver, heart, brain, abdominal cavity, pelviccavity, thoracic cavity, guts and reproductive tissue). In the skin,treatment may change the morphology and organization of collagen fibersand may result in making the scars less visible and blend in with thesurrounding skin. As used herein, prevention, reduction or inhibition ofscarring encompasses any degree of prevention, reduction or inhibitionin scarring as compared to the level of scarring occurring in acontrol-treated or untreated wound.

Prevention, reduction or inhibition of compromised skin, such ascompromised skin associated with dermal scarring, can be assessed and/ormeasured with reference to microscopic and/or macroscopiccharacteristics. Macroscopic characteristics may include color, height,surface texture and stiffness of the skin. In some instances,prevention, reduction or inhibition of compromised skin may bedemonstrated when the color, height, surface texture and stiffness ofthe skin resembles that of normal skin more closely after treatment thandoes a control that is untreated. Microscopic assessment of compromisedskin may involve examining characteristics such as thickness and/ororientation and/or composition of the extracellular matrix (ECM) fibers,and cellularity of the compromised skin. In some instances, prevention,reduction or inhibition of compromised skin may be demonstrated when thethickness and/or orientation and/or composition of the extracellularmatrix (ECM) fibers, and/or cellularity of the compromised skinresembles that of normal skin more closely after treatment than does acontrol that is untreated.

In some aspects, methods associated with the invention are used forcosmetic purposes, at least in part to contribute to improving thecosmetic appearance of compromised skin. In some embodiments, methodsassociated with the invention may be used to prevent, reduce or inhibitcompromised skin such as scarring of wounds covering joints of the body.In other embodiments, methods associated with the invention may be usedto promote accelerated wound healing and/or prevent, reduce or inhibitscarring of wounds at increased risk of forming a contractile scar,and/or of wounds located at sites of high skin tension.

In some embodiments, methods associated with the invention can beapplied to promoting healing of compromised skin in instances wherethere is an increased risk of pathological scar formation, such ashypertrophic scars and keloids, which may have more pronounceddeleterious effects than normal scarring. In some embodiments, methodsdescribed herein for promoting accelerated healing of compromised skinand/or preventing, reducing or inhibiting scarring are applied tocompromised skin produced by surgical revision of pathological scars.

Aspects of the invention can be applied to compromised skin caused byburn injuries. Healing in response to burn injuries can lead to adversescarring, including the formation of hypertrophic scars. Methodsassociated with the invention can be applied to treatment of allinjuries involving damage to an epithelial layer, such as injuries tothe skin in which the epidermis is damaged. Other non-limiting examplesof injuries to epithelial tissue include injuries involving therespiratory epithelia, digestive epithelia or epithelia surroundinginternal tissues or organs.

RNAi to Treat Liver Fibrosis

In some embodiments, methods associated with the invention are used totreat liver fibrosis. Liver fibrosis is the excessive accumulation ofextracellular matrix proteins, including collagen, that occurs in mosttypes of chronic liver diseases. It is the scarring process thatrepresents the liver's response to injury. Advanced liver fibrosisresults in cirrhosis, liver failure, and portal hypertension and oftenrequires liver transplantation. In the same way as skin and other organsheal wounds through deposition of collagen and other matrix constituentsso the liver repairs injury through the deposition of new collagen.Activated hepatic stellate cells, portal fibroblasts, and myofibroblastsof bone marrow origin have been identified as major collagen-producingcells in the injured liver. These cells are activated by fibrogeniccytokines such as TGF-β1, angiotensin II, and leptin. In someembodiments, methods provided herein are aimed at inhibiting theaccumulation of fibrogenic cells and/or preventing the deposition ofextracellular matrix proteins. In some embodiments, RNAi molecules(including sd-rxRNA and rxRNAori) may be designed to target CTGF,TGF-β1, angiotensin II, and/or leptin. In some embodiments, RNAimolecules (including sd-rxRNA and rxRNAori) may be designed to targetthose genes listed in Tables 1-25.

Trabeculectomy Failure

Trabeculectomy is a surgical procedure designed to create a channel orbleb though the sclera to allow excess fluid to drain from the anteriorof the eye, leading to reduced intracocular pressure (IOP), a riskfactor for glaucoma-related vision loss. The most common cause oftrabeculectomy failure is blockage of the bleb by scar tissue. Incertain embodiments, the sd-rxRNA is used to prevent formation of scartissue resulting from a trabeculectomy. In some embodiments, thesd-rxRNA targets connexin 43. In other embodiments, the sd-rxRNA targetsproyly 4-hydroxylase. In yet other embodiments, the sd-rxRNA targetsprocollagen C-protease.

Target Genes

It should be appreciated that based on the RNAi molecules designed anddisclosed herein, one of ordinary skill in the art would be able todesign such RNAi molecules to target a variety of different genesdepending on the context and intended use. For purposes of pre-treating,treating, or preventing compromised skin and/or promoting wound healingand/or preventing, reducing or inhibiting scarring, one of ordinaryskill in the art would appreciate that a variety of suitable targetgenes could be identified based at least in part on the known orpredicted functions of the genes, and/or the known or predictedexpression patterns of the genes. Several non-limiting examples of genesthat could be targeted by RNAi molecules for pre-treating, treating, orpreventing compromised skin and/or promoting wound healing and/orpreventing, reducing or inhibiting scarring include genes that encodefor the following proteins: Transforming growth factor β (TGFβ1, TGFβ2,TGFβ3), Osteopontin (SPP1), Connective tissue growth factor (CTGF),Platelet-derived growth factor (PDGF), Hypoxia inducible factor-1α(HIF1α), Collagen I and/or III, Prolyl 4-hydroxylase (P4H), ProcollagenC-protease (PCP), Matrix metalloproteinase 2, 9 (MMP2, 9), Integrins,Connexin, Histamine H1 receptor, Tissue transglutaminase, Mammaliantarget of rapamycin (mTOR), HoxB13, VEGF, IL-6, SMAD proteins, Ribosomalprotein S6 kinases (RSP6), Cyclooxygenase-2 (COX-2/PTGS2), Cannabinoidreceptors (CB1, CB2), and/or miR29b.

Transforming growth factor β proteins, for which three isoforms exist inmammals (TGFβ1, TGFβ2, TGFβ3), are secreted proteins belonging to asuperfamily of growth factors involved in the regulation of manycellular processes including proliferation, migration, apoptosis,adhesion, differentiation, inflammation, immuno-suppression andexpression of extracellular proteins. These proteins are produced by awide range of cell types including epithelial, endothelial,hematopoietic, neuronal, and connective tissue cells. RepresentativeGenbank accession numbers providing DNA and protein sequence informationfor human TGFβ1, TGFβ2 and TGFβ3 are BT007245, BC096235, and X14149,respectively. Within the TGF family, TGFβ1 and TGFβ2 but not TGFβ3represent suitable targets. The alteration in the ratio of TGFβ variantswill promote better wound healing and will prevent excessive scarformation.

Osteopontin (OPN), also known as Secreted phosphoprotein 1 (SPP1), BoneSinaloprotein 1 (BSP-1), and early T-lymphocyte activation (ETA-1) is asecreted glycoprotein protein that binds to hydroxyapatite. OPN has beenimplicated in a variety of biological processes including boneremodeling, immune functions, chemotaxis, cell activation and apoptosis.Osteopontin is produced by a variety of cell types includingfibroblasts, preosteoblasts, osteoblasts, osteocytes, odontoblasts, bonemarrow cells, hypertrophic chondrocytes, dendritic cells, macrophages,smooth muscle, skeletal muscle myoblasts, endothelial cells, andextraosseous (non-bone) cells in the inner ear, brain, kidney, deciduum,and placenta. Representative Genbank accession number providing DNA andprotein sequence information for human Osteopontin are NM_000582.2 andX13694.

Connective tissue growth factor (CTGF), also known as Hypertrophicchondrocyte-specific protein 24, is a secreted heparin-binding proteinthat has been implicated in wound healing and scleroderma. Connectivetissue growth factor is active in many cell types including fibroblasts,myofibroblasts, endothelial and epithelial cells. Representative Genbankaccession number providing DNA and protein sequence information forhuman CTGF are NM_001901.2 and M92934.

The Platelet-derived growth factor (PDGF) family of proteins, includingseveral isoforms, are secreted mitogens. PDGF proteins are implicated inwound healing, at least in part, because they are released fromplatelets following wounding. Representative Genbank accession numbersproviding DNA and protein sequence information for human PDGF genes andproteins include X03795 (PDGFA), X02811 (PDGFB), AF091434 (PDGFC),AB033832 (PDGFD).

Hypoxia inducible factor-1α (HIF1α), is a transcription factor involvedin cellular response to hypoxia. HIF1α is implicated in cellularprocesses such as embryonic vascularization, tumor angiogenesis andpathophysiology of ischemic disease. A representative Genbank accessionnumber providing DNA and protein sequence information for human HIF1α isU22431.

Collagen proteins are the most abundant mammalian proteins and are foundin tissues such as skin, tendon, vascular, ligature, organs, and bone.Collagen I proteins (such as COL1A1 and COL1A2) are detected in scartissue during wound healing, and are expressed in the skin. Collagen IIIproteins (including COL3A1) are detected in connective tissue in wounds(granulation tissue), and are also expressed in skin. RepresentativeGenbank accession numbers providing DNA and protein sequence informationfor human Collagen proteins include: Z74615 (COL1A1), J03464 (COL1A2)and X14420 (COL3A1).

Prolyl 4-hydroxylase (P4H), is involved in production of collagen and inoxygen sensing. A representative Genbank accession number providing DNAand protein sequence information for human P4H is AY198406.

Procollagen C-protease (PCP) is another target.

Matrix metalloproteinase 2, 9 (MMP2, 9) belong to the metzincinmetalloproteinase superfamily and are zinc-dependent endopeptidases.These proteins are implicated in a variety of cellular processesincluding tissue repair. Representative Genbank accession numbersproviding DNA and protein sequence information for human MMP proteinsare M55593 (MMP2) and J05070 (MMP9).

Integrins are a family of proteins involved in interaction andcommunication between a cell and the extracellular matrix. Vertebratescontain a variety of integrins including α₁β₁, α₂β₁, α₄β₁, α₅β₁, α₆β₁,α_(L)β₂, α_(M)β₂, α_(IIb)β₃, α_(v)β₃, α_(v)β₅, α_(v)β₆, α₆β₄.

Connexins are a family of vertebrate transmembrane proteins that formgap junctions. Several examples of Connexins, with the accompanying genename shown in brackets, include Cx23 (GJE1), Cx25 (GJB7), Cx26 (GJB2),Cx29 (GJE1), Cx30 (GJB6), Cx30.2 (GJC3), Cx30.3 (GJB4), Cx31 (GJB3),Cx31.1 (GJB5), Cx31.9 (GJC1/GJD3), Cx32 (GJB1), Cx33 (GJA6), Cx36(GJD2/GJA9), Cx37 (GJA4), Cx39 (GJD4), Cx40 (GJA5), Cx40.1 (GJD4), Cx43(GJA1), Cx45 (GJC1/GJA7), Cx46 (GJA3), Cx47 (GJC2/GJA12), Cx50 (GJA8),Cx59 (GJA10), and Cx62 (GJA10).

Histamine H1 receptor (HRH1) is a metabotropic G-protein-coupledreceptor involved in the phospholipase C and phosphatidylinositol (PIP2)signaling pathways. A representative Genbank accession number providingDNA and protein sequence information for human HRH1 is Z34897.

Tissue transglutaminase, also called Protein-glutaminegamma-glutamyltransferase 2, is involved in protein crosslinking and isimplicated is biological processes such as apoptosis, cellulardifferentiation and matrix stabilization. A representative Genbankaccession number providing DNA and protein sequence information forhuman Tissue transglutaminase is M55153.

Mammalian target of rapamycin (mTOR), also known asSerine/threonine-protein kinase mTOR and FK506 binding protein12-rapamycin associated protein 1 (FRAP1), is involved in regulatingcell growth and survival, cell motility, transcription and translation.A representative Genbank accession number providing DNA and proteinsequence information for human mTOR is L34075.

HoxB13 belongs to the family of Homeobox proteins and has been linked tofunctions such as cutaneous regeneration and fetal skin development. Arepresentative Genbank accession number providing DNA and proteinsequence information for human HoxB13 is U57052.

Vascular endothelial growth factor (VEGF) proteins are growth factorsthat bind to tyrosine kinase receptors and are implicated in multipledisorders such as cancer, age-related macular degeneration, rheumatoidarthritis and diabetic retinopathy. Members of this protein familyinclude VEGF-A, VEGF-B, VEGF-C and VEGF-D. Representative Genbankaccession numbers providing DNA and protein sequence information forhuman VEGF proteins are M32977 (VEGF-A), U43368 (VEGF-B), X94216(VEGF-C), and D89630 (VEGF-D).

Interleukin-6 (IL-6) is a cytokine involved in stimulating immuneresponse to tissue damage. A representative Genbank accession numberproviding DNA and protein sequence information for human IL-6 is X04430.

SMAD proteins (SMAD1-7, 9) are a family of transcription factorsinvolved in regulation of TGFβ signaling. Representative Genbankaccession numbers providing DNA and protein sequence information forhuman SMAD proteins are U59912 (SMAD1), U59911 (SMAD2), U68019 (SMAD3),U44378 (SMAD4), U59913 (SMAD5), U59914 (SMAD6), AF015261 (SMAD7), andBC011559 (SMAD9).

Ribosomal protein S6 kinases (RSK6) represent a family ofserine/threonine kinases involved in activation of the transcriptionfactor CREB. A representative Genbank accession number providing DNA andprotein sequence information for human Ribosomal protein S6 kinasealpha-6 is AF184965.

Cyclooxygenase-2 (COX-2), also called Prostaglandin G/H synthase 2(PTGS2), is involved in lipid metabolism and biosynthesis of prostanoidsand is implicated in inflammatory disorders such as rheumatoidarthritis. A representative Genbank accession number providing DNA andprotein sequence information for human COX-2 is AY462100.

Cannabinoid receptors, of which there are currently two known subtypes,CB1 and CB2, are a class of cell membrane receptors under the Gprotein-coupled receptor superfamily. The CB1 receptor is expressedmainly in the brain, but is also expressed in the lungs, liver andkidneys, while the CB2 receptor is mainly expressed in the immune systemand in hematopoietic cells. A representative Genbank accession numberproviding DNA and protein sequence information for human CB1 isNM_001160226, NM_001160258, NM_001160259, NM_001160260, NM_016083, andNM_033181.

miR29b (or miR-29b) is a microRNA (miRNA), which is a short (20-24 nt)non-coding RNA involved in post-transcriptional regulation of geneexpression in multicellular organisms by affecting both the stabilityand translation of mRNAs. miRNAs are transcribed by RNA polymerase II aspart of capped and polyadenylated primary transcripts (pri-miRNAs) thatcan be either protein-coding or non-coding. The primary transcript iscleaved by the Drosha ribonuclease III enzyme to produce anapproximately 70-nt stem-loop precursor miRNA (pre-miRNA), which isfurther cleaved by the cytoplasmic Dicer ribonuclease to generate themature miRNA and antisense miRNA star (miRNA*) products. The maturemiRNA is incorporated into a RNA-induced silencing complex (RISC), whichrecognizes target mRNAs through imperfect base pairing with the miRNAand most commonly results in translational inhibition or destabilizationof the target mRNA. A representative miRBase accession number for miR29bis MI0000105 (website:mirbase.org/cgi-bin/mirna_entry.pl?acc=MI0000105).

In some embodiments, the sd-rxRNA targets connexin 43 (CX43). This geneis a member of the connexin gene family. The encoded protein is acomponent of gap junctions, which are composed of arrays ofintercellular channels that provide a route for the diffusion of lowmolecular weight materials from cell to cell. The encoded protein is themajor protein of gap junctions in the heart that are thought to have acrucial role in the synchronized contraction of the heart and inembryonic development. A related intronless pseudogene has been mappedto chromosome 5. Mutations in this gene have been associated withoculodentodigital dysplasia and heart malformations. RepresentativeGenbank accession numbers providing DNA and protein sequence informationfor human CX43 genes and proteins include NM_000165 and NP_000156.

In other embodiments, the sd-rxRNA targets prolyl 4-hydroxylase (P4HTM).The product of this gene belongs to the family of prolyl 4-hydroxylases.This protein is a prolyl hydroxylase that may be involved in thedegradation of hypoxia-inducible transcription factors under normoxia.It plays a role in adaptation to hypoxia and may be related to cellularoxygen sensing. Alternatively spliced variants encoding differentisoforms have been identified. Representative Genbank accession numbersproviding DNA and protein sequence information for human P4HTM genes andproteins include NM_177938, NP_808807, NM_177939, and NP_808808.

In certain embodiments, the sd-rxRNA targets procollagen C-protease.

EXAMPLES Example 1: In Vivo Gene Silencing in Skin after Local Deliveryof Sd-rxRNA

Demonstrated herein is gene silencing in skin following administrationof sd-rxRNA molecules. Rat incision models were used which included 6dorsal incisions per animal. Analysis included monitoring of digitalimages, detection of target gene expression, scar assessment, andhistology. FIG. 1 reveals an expression profile of several genesincluding MAP4K4, SPP1, CTGF, PTGS2 and TGFB1. As expected, whenexpression of these genes was monitored post-incision, target geneexpression was elevated early and then returned to normal by day 10.

FIG. 2 presents an overview of intradermal injection experiments withsd-rxRNA molecules. 6 intradermal injections were performed at eachsite. Each injection consisted of approximately 34μ, 300 μg total.Images were taken before injection and 15 minutes after the firstinjection.

FIG. 3 demonstrates in vivo silencing following intradermal injection ofsd-rxRNA in rats. 6 injections were made per dose. 300 μg in PBS wasinjected on days 1 & 2 (2 doses) or on day 2 (1 dose). 5 incisions siteswere made per treatment. Incisions were 1 cm. 3 mm skin biopsies wereharvested 48 hours after the last dose and target expression wasdetermined by QPCR.

FIG. 4 demonstrates in vivo silencing of MAP4K4, PPIB and CTGFexpression in rats following intradermal injection of sd-rxRNAmolecules. A single intradermal injection of PBS (vehicle), or 300 ug ofMAP4K4, or 2 different CTGF or PPIB targeting sd-rxRNA were injected at6 sites. 3 mm skin biopsies harvested 48 hours post injection andprocessed for RNA. Data was analyzed by QPCR and normalized to B-Actin.PBS was set to 1. Data was graphed as a percent reduction in targetedgene expression relative to non-targeting sd-rxRNA (i.e. targeting othergene). Gene expression from untreated skin samples on treated animalsare similar to PBS treated or sham controls.

FIG. 5 demonstrates in vivo silencing in mice following intradermalinjection of sd-rxRNA molecules. C57BL/6 mice were used, with n=7 whealsites active and PBS. The control group consisted of 12. 300 ug wasadministered in 50 ul/injections. 3 mm biopsies were processed for RNA,and target expression determined by QPCR. Expression was normalized tohousekeeping gene cyclophilin B.

FIG. 6 reveals the in vitro potency and in vivo effectiveness of 2different sd-rxRNAs targeting PPIB. Two PPIB sd-rxRNAs with differentEC50s were compared in vivo. Similar in vivo results were obtained with1 injection of 300 μg

FIGS. 7 and 8 demonstrate the duration of gene silencing achievedthrough administration of sd-rxRNA. There were 6 injection sites peranimal. 3 mm skin biopsies were harvested on days 3, 5, and 8. RNA wasisolated and gene expression was analyzed by qPCR and normalized toB-Actin

FIG. 9 compares two different dosage regimens, Days 1 and 3 vs. Days 0and 2. There were 6 injection sites per animal. 3 mm skin biopsies wereharvested on days 3, 5, and 8. RNA was isolated and gene expression wasanalyzed by qPCR and normalized to B-Actin.

Example 2: Identification of Potent Sd-rxRNAs

Up to 300 rxRNA ori compounds were screened for 5 anti-scarring targets.Optimal sequences in SPP1, CTGF, PTGS2, TGFB1 and TGFB2 for sd-rxRNAdevelopment were identified using a sequence selection algorithm. Thealgorithm selects sequences based on the following criteria: a GCcontent greater than 32% but less than 47%, homology to specific animalmodels (e.g., mouse or rat), avoidance of 5 or more U/U stretches and/or2 or more G/C stretches, an off-target hit score of less than 500, andavoidance of sequences contained within the 5′ UTR.

The sequences were developed initially as 25 nucleotide blunt-endedduplexes with O-methyl modification. Such sequences were screened invarious cell lines to identify those were most efficient in reducinggene expression. Several concentrations of the RNA molecules, such as0.025, 0.1 and 0.25 nM, were tested, and suitable concentrations toscreen for bDNA were determined. A bDNA was then run of a full screen ata desired concentration. Dose response curves were generated todetermine the most potent sequences. Hyperfunctional hits were thosewith an EC50 of less than 100 pM in lipid transfection. Potent moleculeswere selected to be developed into sd-rxRNAs based on the parametersdescribed throughout the application and a secondary screen wasconducted using the sd-rxRNAs.

FIGS. 10-12 reveal that CTGF sd-rxRNAs are efficacious in mediating genesilencing. A dose response for CTGF is indicated in FIG. 12.

FIGS. 13-14 reveal that the original sd-rxrNA screen had a low hit rate.FIG. 15 reveals PTGS2 knockdown using sd-rxRNA against PTGS2. FIGS.16-24 reveal that hTGFB1, TGFB, TGFB2 sd-rxRNAs are capable of mediatinggene silencing. FIGS. 25-28 shows the identification of potent hSPP1sd-rxRNAs.

Example 3: Linker Chemistry

FIG. 36 demonstrates that variation of linker chemistry does notinfluence silencing activity of sd-rxRNAs in vitro. Two different linkerchemistries were evaluated, a hydroxyproline linker and ribo linker, onmultiple sd-rxRNAs (targeting Map4k4 or PPIB) in passive uptake assaysto determine linkers which favor self delivery. HeLa cells weretransfected in the absence of a delivery vehicle (passive transfection)with sd-rxRNAs at 1 uM, 0.1 uM or 0.01 uM for 48 hrs. Use of eitherlinker results in an efficacious delivery of sd-rxRNA.

The ribo linker used in Example 5 had the following structure:

Example 4: Optimization of Target Sequences

Chemical optimization was performed for several lead sequences,including CTGF, PTGS2, TGFβ1, and TGFβ2. Multiples versions of sd-rxRNAleads were synthesized. The sense strand was further O-methyl modified,such as by introduction of O-methyl blocks on the ends, introduction ofO-methyl phosphorothioate blocks at the ends or introduction of fulO-methyl modification with a phosphorothioate block on the 3′ end.

The guide strand was modified to decrease the number of 2′F, substitute2′F with O-methyl, vary the number of ribonucleotides, eliminatestretches of ribonucleotides, minimize the presence of ribonucleotidesnext to the phosphorothioate modifications, and if possible removeribonucleotides from the single stranded region.

Various versions of compounds were synthesized and their efficacy wastested in vitro using passive uptake. The efficacy and toxicity of theoptimized compounds was evaluated in vivo.

All compounds show in vivo efficacy. Initially, activity required highconcentration and at high concentrations some compound demonstratedinjection site reaction. However, data indicated that efficacy andtoxicity in vivo could be dramatically improved by enhancement ofstability and reduction of 2′ F content. In some instances, toxicity, atleast in part, was related to the presence of cholesterol containingshort oligomer metabolites. This type of toxicity is expected to bereduced by stabilization. In general, chemical stabilization was welltolerated. Exact chemical optimization patterns differed for variouscompound. In some cases, complete stabilization resulted in a slightlynegative impact on activity. For most target sites, at least twochemically optimized leads were identified: chemically optimized with invitro efficacy retained or improved compared to an Early Lead and FullyModified, where in vitro efficacy is slightly reduced.

In general, a fully O-methyl modified sense strand is acceptable. Insome instances, it is preferable if less than all of the nucleotides inthe sense strand are O-methyl modified. In some instances, the 3′ end ofthe passenger strand contained a PS/O-METHYL block (2 O-methylmodifications and two 2 phosphorothioate modifications) to insuremaximized stability next to the hydrophobic conjugate.

For all compounds, it was possible to identify functional heavilystabilized leads. In some instances, the number of ribonucleotides percompounds was reduced to 4-6. Multiples versions of sd-rxRNA leads weresynthesized. The number of 2′F modified purines was limited wherepossible to improve manufacturability but some optimized compounds docontain some 2′F modified purines.

Optimized Compounds

A summary of CTGF lead compounds is shown in Table 24. PTGS leads areshown in Table 25. hTGFβ1 leads are shown in Table 26 and hTGFβ2 leadsare shown in Table 27. Lead compounds were tested for in vitro efficacywith varying levels of methylation of the sense strand.

For CTGF Lead 1 (L1), the fully O-methyl modified sense strand wasefficacious having a slight reduction in in vitro efficacy.

For CTGF L2, the fully O-methyl modified sense strand was efficacious,having a slight reduction in in vitro efficacy.

For CTGF L3, the fully O-methyl modified sense strand was partiallyefficacious, having a reduction in in vitro efficacy.

For CTGF L4, the fully O-methyl modified sense strand was partiallyefficacious, having a slight reduction in in vitro efficacy.

For PTGS2 L1 and L2, the fully O-methyl modified sense strand was notefficacious.

For TGFβ1 hL3, the fully O-methyl modified sense strand was efficacious.

For TGFβ2, the fully O-methyl modified sense strand was efficacious.

In Vivo Efficacy of Lead Compounds

The activity of lead compounds was tested in vivo both in cell cultureand in animal models. FIGS. 33 and 34 demonstrate the activity ofoptimized CTGF L1 compounds. FIG. 35 demonstrates the in vitro stabilityof the CTGF L1 compounds. FIGS. 36 and 37 demonstrate the activity ofoptimized CTGF L2 compounds. FIG. 38 demonstrates the in vitro stabilityof the CTGF L2 compounds. FIG. 39 provides a summary of the in vivoactivity of CTGF lead compounds. FIG. 40 demonstrates the efficacy ofCTGF L1 compounds in skin biopsies from rats. FIG. 41 shows the efficacyof CTGF L2 compounds in achieving gene silencing.

FIG. 42 demonstrates CTGF silencing following intradermal injection ofRXi-109. FIG. 43 demonstrates the duration of CTGF silencing in skinafter intradermal injection of the sd-rxRNA in SD rats. Eight millimeterskin biopsies were harvested, and mRNA levels were quantified by QPCRand normalized to a housekeeping gene. Shown is percent (%) silencingvs. Non Targeting Control (NTC); PBS at each time point is oneexperimental group; * p≤0.04; ** p≤0.002.

FIGS. 44-46 show that CTGF L3 and L4 compounds are also active. FIG. 47demonstrates changes in mRNA expression levels of CTGF, α-SM actin,collagen 1A2, and collagen 3A1 after intradermal injection of CTFGsd-rxRNA in SD rats. mRNA levels were quantified by qPCR. Substantialreduction in CTGF expression is observed.

FIG. 49 demonstrates that administration of sd-rxRNAs decreases woundwidth over the course of at least 9 days. The graph shows microscopicmeasurements of wound width in rats on days 3, 6, and 9 post-wounding.Each group represents 5 rats. Two non-serial sections from each woundwere measured and the average width of the two was calculated per wound.*p<0.05 vs. PBS an NTC.

FIG. 50 demonstrates that administration of sd-rxRNAs decreases woundarea over the course of at least 9 days. The graph shows microscopicmeasurements of wound width in rats on days 3, 6, and 9 post-wounding.Each group represents 5 rats. Two non-serial sections from each woundwere measured and the average width of the two was calculated per wound.*p<0.05 vs. PBS an NTC.

FIG. 51 demonstrates that administration of sd-rxRNAs increase thepercentage of wound re-epithelialization over the course of at least 9days. The graph shows microscopic measurements of wound width in rats ondays 3, 6, and 9 post-wounding. Each group represents 5 rats. Twonon-serial sections from each wound were measured and the average widthof the two was calculated per wound. *p<0.05 vs. PBS an NTC.

FIG. 52 demonstrates that administration of sd-rxRNAs increases theaverage granulation tissue maturity scores over the course of at least 9days. The graph shows microscopic measurements of wound width in rats ondays 3, 6, and 9 post-wounding (5=mature, 1=immature). Each grouprepresents 5 rats. FIG. 53 demonstrates CD68 labeling in day 9 wounds(0=no labeling, 3=substantial labeling). Each group represents 5 rats.

FIG. 54 demonstrates that CTGF leads have different toxicity levels invitro. FIG. 55 shows percentage (%) of cell viability after RXi-109 doseescalation (oligos formulated in PBS).

FIG. 56 is a schematic of a non-limiting example of a Phase 1 and 2clinical trial design for lead compounds. This schematic represents adivided dose, single day ascending dose clinical trial.

FIG. 57 is a schematic of a non-limiting example of a Phase 1 and 2clinical trial design. This schematic represents a divided dose,multi-day ascending dose clinical trial.

Activity of PTGS2, TGFβ1 and TGFβ2 leads was also tested. FIGS. 59 and60 demonstrate activity of PTGS2 L1 and L2 compounds. FIGS. 61 and 62demonstrate the activity of h TGFβ1 compounds and FIGS. 63 and 64demonstrate the activity of hTGFβ2 compounds.

Gene knock-down in liver was also tested following tail vein injectionmice. FIG. 58 demonstrates a percent (%) decrease in PPIB expression inthe liver relative to PBS control. Lipoid formulated rxRNAs (10 mg/kg)were delivery systemically to Balb/c mice (n=5) by single tail veininjections. Liver tissue was harvested at 24 hours after injection andexpression was analyzed by qPCR (normalized to β-actin). Map4K4 rxRNAorialso showed significant silencing (˜83%, p<0.001) although Map4K4sd-rxRNA did not significantly reduce target gene expression (˜17%,p=0.019). TD.035.2278, Published lipidoid delivery reagent, 98N12-5(1),from Akinc, 2009.

Table 1 provides sequences tested in the Original sd-rxRNA screen.

Table 2 demonstrates inhibition of gene expression with PTGS2 orisequences.

Table 3 demonstrates non-limiting examples of PTGS2 sd-rxRNA sequences.

Table 4 demonstrates non-limiting examples of TGFB1 sd-rxRNA sequences.

Table 5 demonstrates inhibition of gene expression with hTGFB1 orisequences.

Table 6 demonstrates inhibition of gene expression with hTGFB2 orisequences.

Table 7 demonstrates non-limiting examples of hTGFB2 sd-rxRNA sequences.

Table 8 demonstrates non-limiting examples of hSPP1 sd-rxRNA sequences.

Table 9 demonstrates inhibition of gene expression with hSPP1 orisequences.

Table 10 demonstrates non-limiting examples of hCTGF sd-rxRNA sequences.

Table 11 demonstrates inhibition of gene expression with hCTGF orisequences.

Table 12 demonstrates inhibition of gene expression with CTGF orisequences.

Table 13 demonstrates inhibition of gene expression with SPP1 sd-rxRNAsequences.

Table 14 demonstrates inhibition of gene expression with PTGS2 sd-rxRNAsequences.

Table 15 demonstrates inhibition of gene expression with CTGF sd-rxRNAsequences.

Table 16 demonstrates inhibition of gene expression with TGFB2 sd-rxRNAsequences.

Table 17 demonstrates inhibition of gene expression with TGFB1 sd-rxRNAsequences.

Table 18 demonstrates inhibition of gene expression with SPP1 sd-rxRNAsequences.

Table 19 demonstrates inhibition of gene expression with PTGS2 sd-rxRNAsequences.

Table 20 demonstrates inhibition of gene expression with CTGF sd-rxRNAsequences.

Table 21 demonstrates inhibition of gene expression with TGFB2 sd-rxRNAsequences.

Table 22 demonstrates inhibition of gene expression with TGFB1 sd-rxRNAsequences.

Table 23 provides non-limiting examples of CB1 sequences.

Table 24 provides a summary of CTGF Leads.

Table 25 provides a summary of PTGS2 Leads.

Table 26 provides a summary of TGFβ1 Leads.

Table 27 provides a summary of TGFβ1 Leads.

TABLE 1 Original sd-rxRNA screen Oligo SEQ ID SEQ ID ID# G1- NOSense-sd-rxRNA GII NO AS-sd-rxRNA-GII 14394 TGFB1 1 GmCmUAAmUGGmUGGAA- 25′-P-mU(2′-F-U)(2′-F-C)(2′-F-C)A(2′- chol F-C)(2′-F-C)A(2′-F-U)(2′-F-U)AGmC*A*mC*G*mC*G*G 14395 TGFB1 3 mUGAmUmCGmUGmCGmC 45′-P-mGAG(2′-F-C)G(2′-F-C)A(2′-F- mUmC-chol C)GA(2′-F-U)mCA*mU*G*mU*mU*G*G 14396 TGFB1 5 mCAAmUmUmCmCmUGGmC 65′-P-mU(2′-F-C)G(2′-F-C)(2′-F- GA-chol C)AGGAA(2′-F-U)mUG*mU*mU*G*mC*mU*G 14397 TGFB1 7 AGmUGGAmUmCmCAmCGA- 85′-P-mU(2′-F-C)G(2′-F-U)GGA(2′-F- chol U)(2′-F-C)(2′-F-C)AmCmU*mU*mC*mC*A*G*C 14398 TGFB1 9 mUAmCAGmCAAGGmUmCm 105′-P-mGGA(2′-F-C)(2′-F-C)(2′-F- C-chol U)(2′-F-U)G(2′-F-C)(2′-F-U)GmUA*mC*mU*G*mC*G*U 14399 TGFB1 11 AAmCAmUGAmUmCGmUGm 125′-P-mG(2′-F-C)A(2′-F-C)GA(2′-F- C-chol U)(2′-F-C)A(2′-F-U)GmUmU*G*G*A*mC*A*G 14400 TGFB1 13 AmCAmUGAmUmCGmUGmC 145′-P-mCG(2′-F-C)A(2′-F-C)GA(2′-F- G-chol U)(2′-F-C)A(2′-F-U)GmU*mU*G*G*A*mC*A 14401 TGFB1 15 mCAGmCAAGGmUmCmCmU 165′-P-mCAGGA(2′-F-C)(2′-F-C)(2′-F- G-chol U)(2′-F-U)G(2′-F-C)mUG*mU*A*mC*mU*G*C 14402 TGFB1 17 mCmCAAmCAmUGAmUmCG 185′-P-mA(2′-F-C)GA(2′-F-U)(2′-F- mU-chol C)A(2′-F-U)G(2′-F-U)(2′-F-U)GG*A*mC*A*G*mC*U 14403 TGFB1 19 AGmCGGAAGmCGmCAmU- 205′-P-mA(2′-F-U)G(2′-F-C)G(2′-F- chol C)(2′-F-U)(2′-F-U)(2′-F-C)(2′-F-C)GmCmU*mU*mC*A*mC*mC*A 14404 TGFB1 21 GmCAmUmCGAGGmCmCAm 225′-P-mA(2′-F-U)GG(2′-F-C)(2′-F- U-chol C)(2′-F-U)(2′-F-C)GA(2′-F-U)GmC*G*mC*mU*mU*mC*C 14405 TGFB1 23 GAmCmUAmUmCGAmCAmU 245′-P-mCA(2′-F-U)G(2′-F-U)(2′-F- G-chol C)GA(2′-F-U)AGmUmC*mU*mU*G*mC*A*G 14406 TGFB1 25 AmCmCmUGmCAAGAmCmU 265′-P-mUAG(2′-F-U)(2′-F-C)(2′-F- A-chol U)(2′-F-U)G(2′-F-C)AGGmU*G*G*A*mU*A*G 14407 TGFB1 27 GmCmUmCmCAmCGGAGAA- 285′-P-mU(2′-F-U)(2′-F-C)(2′-F-U)(2′-F- chol C)(2′-F-C)G(2′-F-U)GGAGmC*mU*G*A*A*G*C 14408 TGFB2 29 GGmCmUmCmUmCmCmUm 30 5′-P-mU(2′-F-UmCGA-chol C)GAAGGAGAGmCmC*A*mU*mU* mC*G*C 14409 TGFB2 31GAmCAGGAAmCmCmUGG- 32 5′-P-mC(2′-F-C)AGG(2′-F-U)(2′-F- cholU)(2′-F-C)(2′-F-C)(2′-F- U)GmUmC*mU*mU*mU*A*mU*G 14410 TGFB2 33mCmCAAGGAGGmUmUmUA- 34 5′-P-mUAAA(2′-F-C)(2′-F-C)(2′-F- cholU)(2′-F-C)(2′-F-C)(2′-F-U)(2′-F- U)GG*mC*G*mU*A*G*U 14411 TGFB2 35AmUmUmUmCmCAmUmCm 36 5′-P-mUG(2′-F-U)AGA(2′-F- UAmCA-cholU)GGAAAmU*mC*A*mC*mC*mU*C 14412 TGFB2 37 mUmCmCAmUmCmUAmCAA 385′-P-mUG(2′-F-U)(2′-F-U)G(2′-F- mCA-chol U)AGA(2′-F- U)GGA*A*A*mU*mC*A*C14413 TGFB2 39 mUmUmUmCmCAmUmCmU 40 5′-P-mU(2′-F-U)G(2′-F-U)AGA(2′-F-AmCAA-chol U)GGAAA*mU*mC*A*mC*mC*U 14414 TGFB2 41 mCGmCmCAAGGAGGmUmU- 425′-P-mAA(2′-F-C)(2′-F-C)(2′-F-U)(2′- chol F-C)(2′-F-C)(2′-F-U)(2′-F-U)GGmCG*mU*A*G*mU*A*C 14415 TGFB2 43 GmUGGmUGAmUmCAGAA- 445′-P-mU(2′-F-U)(2′-F-C)(2′-F- chol U)GA(2′-F-U)(2′-F-C)A(2′-F-C)(2′-F-C)AmC*mU*G*G*mU*A*U 14416 TGFB2 45 mCmUmCmCmUGmCmUAA 465′-P-mA(2′-F-C)A(2′-F-U)(2′-F- mUGmU-chol U)AG(2′-F-C)AGGAG*A*mU*G*mU*G*G 14417 TGFB2 47 AmCmCmUmCmCAmCAmUA 485′-P-mUA(2′-F-U)A(2′-F-U)G(2′-F- mUA-chol U)GGAGGmU*G*mC*mC*A*mU*C 14418TGFB2 49 AAGmUmCmCAmCmUAGGA- 50 5′-P-mU(2′-F-C)(2′-F-C)(2′-F- cholU)AG(2′-F-U)GGA(2′-F- C)mUmU*mU*A*mU*A*G*U 14419 TGFB2 51mUGGmUGAmUmCAGAAA- 52 5′-P-mU(2′-F-U)(2′-F-U)(2′-F-C)(2′-F- cholU)GA(2′-F-U)(2′-F-C)A(2′-F- C)mCA*mC*mU*G*G*mU*A 14420 TGFB2 53AGmUmCmCAmCmUAGGAA- 54 5′-P-mU(2′-F-U)(2′-F-C)(2′-F-C)(2′-F- cholU)AG(2′-F- U)GGAmCmU*mU*mU*A*mU*A*G 14421 TGFB2 55 AmCGmCmCAAGGAGGmU- 565′-P-mA(2′-F-C)(2′-F-C)(2′-F-U)(2′-F- cholC)(2′-F-C)(2′-F-U)(2′-F-U)GG(2′-F- C)GmU*A*G*mU*A*mC*U 14422 PTGS2 57mCAmCAmUmUmUGAmUm 58 5′-P-mU(2′-F-C)AA(2′-F-U)(2′-F- UGA-cholC)AAA(2′-F- U)GmUG*A*mU*mC*mU*G*G 14423 PTGS2 59 mCAmCmUGmCmCmUmCAA 605′-P-mAA(2′-F-U)(2′-F-U)GAGG(2′-F- mUmU-chol C)AGmUG*mU*mU*G*A*mU*G14424 PTGS2 61 AAAmUAmCmCAGmUmCmU 62 5′-P-mAAGA(2′-F-C)(2′-F-U)GG(2′-F-mU-chol U)A(2′-F- U)mUmU*mC*A*mU*mC*mU*G 14425 PTGS2 63mCAmUmUmUGAmUmUGA 64 5′-P-mUG(2′-F-U)(2′-F-C)AA(2′-F- mCA-chol U)(2′-F-C)AAAmUG*mU*G*A*mU*mC*U

TABLE 2 Inhibition of gene expression with PTGS2 ori sequences TargetGene Gene SEQ ID PTGS2 NM_000963.2 Duplex ID Region Ref Pos No SenseSequence % Expression (0.1 nM) 15147 CDS 176 65CCUGGCGCUCAGCCAUACAGCAAAA 59% 15148 CDS 177 66 CUGGCGCUCAGCCAUACAGCAAAUA72% 15149 CDS 178 67 UGGCGCUCAGCCAUACAGCAAAUCA 77% 15150 CDS 180 68GCGCUCAGCCAUACAGCAAAUCCUA 70% 15151 CDS 182 69 GCUCAGCCAUACAGCAAAUCCUUGA76% 15152 CDS 183 70 CUCAGCCAUACAGCAAAUCCUUGCA 74% 15153 CDS 184 71UCAGCCAUACAGCAAAUCCUUGCUA 47% 15154 CDS 186 72 AGCCAUACAGCAAAUCCUUGCUGUA55% 15155 CDS 187 73 GCCAUACAGCAAAUCCUUGCUGUUA 41% 15156 CDS 188 74CCAUACAGCAAAUCCUUGCUGUUCA 46% 15157 CDS 212 75 CCACCCAUGUCAAAACCGAGGUGUA31% 15158 CDS 243 76 AGUGUGGGAUUUGACCAGUAUAA 23% GA 15159 CDS 244 77GUGUGGGAUUUGACCAGUAUAAG 24% UA 15160 CDS 245 78 UGUGGGAUUUGACCAGUAUAAGU38% GA 15161 CDS 252 79 UUUGACCAGUAUAAGUGCGAUUG 29% UA 15162 CDS 285 80GGAUUCUAUGGAGAAAACUGCUCAA 16% 15163 CDS 337 81 UAUUUCUGAAACCCACUCCAAACAA32% 15164 CDS 338 82 AUUUCUGAAACCCACUCCAAACACA 21% 15165 CDS 339 83UUUCUGAAACCCACUCCAAACACAA 21% 15166 CDS 340 84 UUCUGAAACCCACUCCAAACACAGA45% 15167 CDS 345 85 AAACCCACUCCAAACACAGUGCACA 87% 15168 CDS 347 86ACCCACUCCAAACACAGUGCACUAA 83% 15169 CDS 349 87 CCACUCCAAACACAGUGCACUACAA51% 15170 CDS 350 88 CACUCCAAACACAGUGCACUACAUA 31% 15171 CDS 394 89UUUGGAACGUUGUGAAUAACAUU 43% CA 15172 CDS 406 90UGAAUAACAUUCCCUUCCUUCGAAA 21% 15173 CDS 407 91 GAAUAACAUUCCCUUCCUUCGAAAA32% 15174 CDS 408 92 AAUAACAUUCCCUUCCUUCGAAAUA 27% 15175 CDS 435 93AUUAUGAGUUAUGUGUUGACAUC 27% CA 15176 CDS 437 94 UAUGAGUUAUGUGUUGACAUCCA21% GA 15177 CDS 439 95 UGAGUUAUGUGUUGACAUCCAGA 30% UA 15178 CDS 440 96GAGUUAUGUGUUGACAUCCAGAU 68% CA 15179 CDS 441 97AGUUAUGUGUUGACAUCCAGAUCAA 35% 15180 CDS 442 98 GUUAUGUGUUGACAUCCAGAUCACA36% 15181 CDS 443 99 UUAUGUGUUGACAUCCAGAUCACAA 51% 15182 CDS 444 100UAUGUGUUGACAUCCAGAUCACAUA 24% 15183 CDS 445 101AUGUGUUGACAUCCAGAUCACAUUA 37% 15184 CDS 446 102UGUGUUGACAUCCAGAUCACAUUUA 42% 15185 CDS 448 103UGUUGACAUCCAGAUCACAUUUGAA 18% 15186 CDS 449 104GUUGACAUCCAGAUCACAUUUGAUA 24% 15187 CDS 450 105UUGACAUCCAGAUCACAUUUGAUUA 25% 15188 CDS 452 106GACAUCCAGAUCACAUUUGAUUGAA 27% 15189 CDS 454 107CAUCCAGAUCACAUUUGAUUGACAA 27% 15190 CDS 455 108AUCCAGAUCACAUUUGAUUGACAGA 32% 15191 CDS 456 109UCCAGAUCACAUUUGAUUGACAGUA 40% 15192 CDS 457 110CCAGAUCACAUUUGAUUGACAGUCA 52% 15193 CDS 460 111GAUCACAUUUGAUUGACAGUCCACA 40% 15194 CDS 461 112AUCACAUUUGAUUGACAGUCCACCA 46% 15195 CDS 462 113UCACAUUUGAUUGACAGUCCACCAA 34% 15196 CDS 463 114CACAUUUGAUUGACAGUCCACCAAA 30% 15197 CDS 464 115ACAUUUGAUUGACAGUCCACCAACA 32% 15198 CDS 465 116CAUUUGAUUGACAGUCCACCAACUA 44% 15199 CDS 467 117UUUGAUUGACAGUCCACCAACUUAA 17% 15200 CDS 468 118UUGAUUGACAGUCCACCAACUUACA 22% 15201 CDS 469 119UGAUUGACAGUCCACCAACUUACAA 27% 15202 CDS 470 120GAUUGACAGUCCACCAACUUACAAA 41% 15203 CDS 471 121AUUGACAGUCCACCAACUUACAAUA 39% 15204 CDS 472 122UUGACAGUCCACCAACUUACAAUGA 61% 15205 CDS 473 123UGACAGUCCACCAACUUACAAUGCA 48% 15206 CDS 479 124UCCACCAACUUACAAUGCUGACUAA 29% 15207 CDS 486 125ACUUACAAUGCUGACUAUGGCUACA 35% 15208 CDS 488 126UUACAAUGCUGACUAUGGCUACAAA 32% 15209 CDS 517 127GGGAAGCCUUCUCUAACCUCUCCUA 39% 15210 CDS 518 128GGAAGCCUUCUCUAACCUCUCCUAA 48% 15211 CDS 519 129GAAGCCUUCUCUAACCUCUCCUAUA 19% 15212 CDS 520 130AAGCCUUCUCUAACCUCUCCUAUUA 17% 15213 CDS 524 131CUUCUCUAACCUCUCCUAUUAUACA 17% 15214 CDS 525 132UUCUCUAACCUCUCCUAUUAUACUA 34% 15215 CDS 526 133UCUCUAACCUCUCCUAUUAUACUAA 49% 15216 CDS 601 134GUAAAAAGCAGCUUCCUGAUUCAAA 35% 15217 CDS 602 135UAAAAAGCAGCUUCCUGAUUCAAAA 25% 15218 CDS 606 136AAGCAGCUUCCUGAUUCAAAUGAGA 27% 15219 CDS 615 137CCUGAUUCAAAUGAGAUUGUGGAAA 37% 15220 CDS 616 138 CUGAUUCAAAUGAGAUUGUGGAA27% AA 15221 CDS 636 139 GAAAAAUUGCUUCUAAGAAGAAAGA 37% 15222 CDS 637 140AAAAAUUGCUUCUAAGAAGAAAGUA 56% 15223 CDS 638 141AAAAUUGCUUCUAAGAAGAAAGUUA 25% 15224 CDS 639 142AAAUUGCUUCUAAGAAGAAAGUUCA 34% 15225 CDS 678 143GGCUCAAACAUGAUGUUUGCAUUCA 68% 15226 CDS 679 144GCUCAAACAUGAUGUUUGCAUUCUA 51% 15227 CDS 680 145CUCAAACAUGAUGUUUGCAUUCUUA 50% 15228 CDS 682 146 CAAACAUGAUGUUUGCAUUCUUU51% GA 15229 CDS 683 147 AAACAUGAUGUUUGCAUUCUUUG 63% CA 15230 CDS 722148 UCAGUUUUUCAAGACAGAUCAUAAA 45% 15231 CDS 723 149CAGUUUUUCAAGACAGAUCAUAAGA 59% 15232 CDS 724 150AGUUUUUCAAGACAGAUCAUAAGCA 80% 15233 CDS 725 151GUUUUUCAAGACAGAUCAUAAGCGA 55% 15234 CDS 726 152UUUUUCAAGACAGAUCAUAAGCGAA 53% 15235 CDS 776 153CCAUGGGGUGGACUUAAAUCAUAUA 56% 15236 CDS 787 154ACUUAAAUCAUAUUUACGGUGAAAA 63% 15237 CDS 788 155CUUAAAUCAUAUUUACGGUGAAACA 43% 15238 CDS 789 156UUAAAUCAUAUUUACGGUGAAACUA 48% 15239 CDS 790 157UAAAUCAUAUUUACGGUGAAACUCA 56% 15240 CDS 792 158AAUCAUAUUUACGGUGAAACUCUGA 46% 15241 CDS 793 159AUCAUAUUUACGGUGAAACUCUGGA 64% 15242 CDS 799 160UUUACGGUGAAACUCUGGCUAGACA 35% 15243 CDS 819 161AGACAGCGUAAACUGCGCCUUUUCA 65% 15244 CDS 820 162GACAGCGUAAACUGCGCCUUUUCAA 51% 15245 CDS 821 163ACAGCGUAAACUGCGCCUUUUCAAA 48% 15246 CDS 822 164CAGCGUAAACUGCGCCUUUUCAAGA 61% 15247 CDS 823 165AGCGUAAACUGCGCCUUUUCAAGGA 48% 15248 CDS 828 166AAACUGCGCCUUUUCAAGGAUGGAA 42% 15249 CDS 830 167ACUGCGCCUUUUCAAGGAUGGAAAA 29% 15250 CDS 861 168 UAUCAGAUAAUUGAUGGAGAGAU32% GA 15251 CDS 862 169 AUCAGAUAAUUGAUGGAGAGAUG 55% UA 15252 CDS 863170 UCAGAUAAUUGAUGGAGAGAUGU 50% AA 15253 CDS 864 171CAGAUAAUUGAUGGAGAGAUGUA 50% UA 15254 CDS 865 172 AGAUAAUUGAUGGAGAGAUGUAU55% CA 15255 CDS 866 173 GAUAAUUGAUGGAGAGAUGUAUC 65% CA 15256 CDS 867174 AUAAUUGAUGGAGAGAUGUAUCC 54% UA 15257 CDS 880 175AGAUGUAUCCUCCCACAGUCAAAGA 78% 15258 CDS 881 176GAUGUAUCCUCCCACAGUCAAAGAA 79% 15259 CDS 882 177AUGUAUCCUCCCACAGUCAAAGAUA 49% 15260 CDS 883 178UGUAUCCUCCCACAGUCAAAGAUAA 28% 15261 CDS 884 179GUAUCCUCCCACAGUCAAAGAUACA 56% 15262 CDS 885 180UAUCCUCCCACAGUCAAAGAUACUA 42% 15263 CDS 887 181UCCUCCCACAGUCAAAGAUACUCAA 45% 15264 CDS 888 182CCUCCCACAGUCAAAGAUACUCAGA 73% 15265 CDS 889 183CUCCCACAGUCAAAGAUACUCAGGA 56% 15266 CDS 891 184CCCACAGUCAAAGAUACUCAGGCAA 80% 15267 CDS 901 185AAGAUACUCAGGCAGAGAUGAUCUA 59% 15268 CDS 935 186AGUCCCUGAGCAUCUACGGUUUGCA 83% 15269 CDS 980 187 UCUGGUGCCUGGUCUGAUGAUGU55% AA 15270 CDS 981 188 CUGGUGCCUGGUCUGAUGAUGUA 56% UA 15271 CDS 982189 UGGUGCCUGGUCUGAUGAUGUAU 43% GA 15272 CDS 983 190GGUGCCUGGUCUGAUGAUGUAUG 41% CA 15273 CDS 984 191 GUGCCUGGUCUGAUGAUGUAUGC42% CA 15274 CDS 985 192 UGCCUGGUCUGAUGAUGUAUGCC 37% AA 15275 CDS 986193 GCCUGGUCUGAUGAUGUAUGCCACA 61% 15276 CDS 1016 194GCUGCGGGAACACAACAGAGUAUGA 44% 15277 CDS 1019 195GCGGGAACACAACAGAGUAUGCGAA 33% 15278 CDS 1038 196UGCGAUGUGCUUAAACAGGAGCAUA 53% 15279 CDS 1039 197GCGAUGUGCUUAAACAGGAGCAUCA 109% 15280 CDS 1040 198CGAUGUGCUUAAACAGGAGCAUCCA 77% 15281 CDS 1042 199AUGUGCUUAAACAGGAGCAUCCUGA 69% 15282 CDS 1043 200UGUGCUUAAACAGGAGCAUCCUGAA 76% 15283 CDS 1044 201GUGCUUAAACAGGAGCAUCCUGAAA 65% 15284 CDS 1045 202UGCUUAAACAGGAGCAUCCUGAAUA 64% 15285 CDS 1084 203UGUUCCAGACAAGCAGGCUAAUACA 41% 15286 CDS 1093 204CAAGCAGGCUAAUACUGAUAGGAGA 24% 15287 CDS 1095 205AGCAGGCUAAUACUGAUAGGAGAGA 50% 15288 CDS 1096 206GCAGGCUAAUACUGAUAGGAGAGAA 51% 15289 CDS 1124 207 UAAGAUUGUGAUUGAAGAUUAUG35% UA 15290 CDS 1125 208 AAGAUUGUGAUUGAAGAUUAUGU 34% GA 15291 CDS 1126209 AGAUUGUGAUUGAAGAUUAUGUG 59% CA 15292 CDS 1127 210GAUUGUGAUUGAAGAUUAUGUGC 41% AA 15293 CDS 1128 211AUUGUGAUUGAAGAUUAUGUGCA 51% AA 15294 CDS 1129 212UUGUGAUUGAAGAUUAUGUGCAA 45% CA 15295 CDS 1131 213GUGAUUGAAGAUUAUGUGCAACACA 37% 15296 CDS 1132 214UGAUUGAAGAUUAUGUGCAACACUA 34% 15297 CDS 1134 215AUUGAAGAUUAUGUGCAACACUUGA 24% 15298 CDS 1136 216UGAAGAUUAUGUGCAACACUUGAGA 37% 15299 CDS 1138 217AAGAUUAUGUGCAACACUUGAGUGA 44% 15300 CDS 1145 218UGUGCAACACUUGAGUGGCUAUCAA 29% 15301 CDS 1149 219CAACACUUGAGUGGCUAUCACUUCA 33% 15302 CDS 1179 220AAAUUUGACCCAGAACUACUUUUCA 35% 15303 CDS 1180 221AAUUUGACCCAGAACUACUUUUCAA 41% 15304 CDS 1181 222AUUUGACCCAGAACUACUUUUCAAA 40% 15305 CDS 1200 223UUCAACAAACAAUUCCAGUACCAAA 49% 15306 CDS 1211 224AUUCCAGUACCAAAAUCGUAUUGCA 27% 15307 CDS 1217 225GUACCAAAAUCGUAUUGCUGCUGAA 31% 15308 CDS 1270 226UUCUGCCUGACACCUUUCAAAUUCA 35% 15309 CDS 1280 227CACCUUUCAAAUUCAUGACCAGAAA 57% 15310 CDS 1284 228UUUCAAAUUCAUGACCAGAAAUACA 42% 15311 CDS 1289 229AAUUCAUGACCAGAAAUACAACUAA 52% 15312 CDS 1327 230ACAACAACUCUAUAUUGCUGGAACA 58% 15313 CDS 1352 231 UGGAAUUACCCAGUUUGUUGAAU35% CA 15314 CDS 1356 232 AUUACCCAGUUUGUUGAAUCAUUCA 41% 15315 CDS 1357233 UUACCCAGUUUGUUGAAUCAUUCAA 58% 15316 CDS 1359 234ACCCAGUUUGUUGAAUCAUUCACCA 52% 15317 CDS 1360 235CCCAGUUUGUUGAAUCAUUCACCAA 66% 15318 CDS 1361 236CCAGUUUGUUGAAUCAUUCACCAGA 54% 15319 CDS 1365 237UUUGUUGAAUCAUUCACCAGGCAAA 47% 15320 CDS 1462 238AGAGCAGGCAGAUGAAAUACCAGUA 65% 15321 CDS 1463 239GAGCAGGCAGAUGAAAUACCAGUCA 66% 15322 CDS 1465 240GCAGGCAGAUGAAAUACCAGUCUUA 22% 15323 CDS 1466 241CAGGCAGAUGAAAUACCAGUCUUUA 43% 15324 CDS 1472 242GAUGAAAUACCAGUCUUUUAAUGAA 23% 15325 CDS 1473 243AUGAAAUACCAGUCUUUUAAUGAGA 61% 15326 CDS 1474 244 UGAAAUACCAGUCUUUUAAUGAG49% UA 15327 CDS 1475 245 GAAAUACCAGUCUUUUAAUGAGUAA 76% 15328 CDS 1476246 AAAUACCAGUCUUUUAAUGAGUACA 51% 15329 CDS 1477 247AAUACCAGUCUUUUAAUGAGUACCA 72% 15330 CDS 1478 248AUACCAGUCUUUUAAUGAGUACCGA 40% 15331 CDS 1479 249UACCAGUCUUUUAAUGAGUACCGCA 53% 15332 CDS 1480 250ACCAGUCUUUUAAUGAGUACCGCAA 39% 15333 CDS 1481 251CCAGUCUUUUAAUGAGUACCGCAAA 41% 15334 CDS 1483 252AGUCUUUUAAUGAGUACCGCAAACA 38% 15335 CDS 1485 253UCUUUUAAUGAGUACCGCAAACGCA 55% 15336 CDS 1486 254CUUUUAAUGAGUACCGCAAACGCUA 63% 15337 CDS 1487 255UUUUAAUGAGUACCGCAAACGCUUA 52% 15338 CDS 1495 256AGUACCGCAAACGCUUUAUGCUGAA 49% 15339 CDS 1524 257UAUGAAUCAUUUGAAGAACUUACAA 65% 15340 CDS 1525 258AUGAAUCAUUUGAAGAACUUACAGA 63% 15341 CDS 1527 259GAAUCAUUUGAAGAACUUACAGGAA 65% 15342 CDS 1529 260AUCAUUUGAAGAACUUACAGGAGAA 43% 15343 CDS 1531 261CAUUUGAAGAACUUACAGGAGAAAA 63% 15344 CDS 1532 262AUUUGAAGAACUUACAGGAGAAAAA 33% 15345 CDS 1574 263GGAAGCACUCUAUGGUGACAUCGAA 62% 15346 CDS 1609 264UGUAUCCUGCCCUUCUGGUAGAAAA 36% 15347 CDS 1614 265CCUGCCCUUCUGGUAGAAAAGCCUA 58% 15348 CDS 1650 266AUCUUUGGUGAAACCAUGGUAGAAA 60% 15349 CDS 1666 267UGGUAGAAGUUGGAGCACCAUUCUA 88% 15350 CDS 1669 268UAGAAGUUGGAGCACCAUUCUCCUA 85% 15351 CDS 1672 269AAGUUGGAGCACCAUUCUCCUUGAA 83% 15352 CDS 1675 270UUGGAGCACCAUUCUCCUUGAAAGA 85% 15353 CDS 1676 271UGGAGCACCAUUCUCCUUGAAAGGA 83% 15354 CDS 1677 272GGAGCACCAUUCUCCUUGAAAGGAA 74% 15355 CDS 1678 273GAGCACCAUUCUCCUUGAAAGGACA 81% 15356 CDS 1679 274AGCACCAUUCUCCUUGAAAGGACUA 86% 15357 CDS 1680 275GCACCAUUCUCCUUGAAAGGACUUA 98% 15358 CDS 1681 276CACCAUUCUCCUUGAAAGGACUUAA 78% 15359 CDS 1682 277ACCAUUCUCCUUGAAAGGACUUAUA 88% 15360 CDS 1683 278CCAUUCUCCUUGAAAGGACUUAUGA 88% 15361 CDS 1762 279UGGGUUUUCAAAUCAUCAACACUGA 78% 15362 CDS 1763 280GGGUUUUCAAAUCAUCAACACUGCA 92% 15363 CDS 1767 281UUUCAAAUCAUCAACACUGCCUCAA 85% 15364 CDS 1770 282CAAAUCAUCAACACUGCCUCAAUUA 84% 15365 CDS 1773 283AUCAUCAACACUGCCUCAAUUCAGA 86% 15366 CDS 1774 284UCAUCAACACUGCCUCAAUUCAGUA 94% 15367 CDS 1775 285CAUCAACACUGCCUCAAUUCAGUCA 84% 15368 CDS 1776 286AUCAACACUGCCUCAAUUCAGUCUA 84% 15369 CDS 1777 287UCAACACUGCCUCAAUUCAGUCUCA 68% 15370 CDS 1778 288CAACACUGCCUCAAUUCAGUCUCUA 73% 15371 CDS 1779 289AACACUGCCUCAAUUCAGUCUCUCA 79% 15372 CDS 1780 290ACACUGCCUCAAUUCAGUCUCUCAA 78% 15373 CDS 1781 291CACUGCCUCAAUUCAGUCUCUCAUA 92% 15374 CDS 1782 292ACUGCCUCAAUUCAGUCUCUCAUCA 89% 15375 CDS 1783 293CUGCCUCAAUUCAGUCUCUCAUCUA 95% 15376 CDS 1784 294UGCCUCAAUUCAGUCUCUCAUCUGA 83% 15377 CDS 1785 295GCCUCAAUUCAGUCUCUCAUCUGCA 46% 15378 CDS 1786 296CCUCAAUUCAGUCUCUCAUCUGCAA 51% 15379 CDS 1787 297CUCAAUUCAGUCUCUCAUCUGCAAA 61% 15380 CDS 1790 298AAUUCAGUCUCUCAUCUGCAAUAAA 30% 15381 CDS 1791 299AUUCAGUCUCUCAUCUGCAAUAACA 32% 15382 CDS 1792 300UUCAGUCUCUCAUCUGCAAUAACGA 30% 15383 CDS 1793 301UCAGUCUCUCAUCUGCAAUAACGUA 38% 15384 CDS 1794 302CAGUCUCUCAUCUGCAAUAACGUGA 67% 15385 CDS 1795 303AGUCUCUCAUCUGCAAUAACGUGAA 71% 15386 CDS 1796 304GUCUCUCAUCUGCAAUAACGUGAAA 81% 15387 CDS 1856 305AGAGCUCAUUAAAACAGUCACCAUA 33% 15388 CDS 1857 306GAGCUCAUUAAAACAGUCACCAUCA 55% 15389 CDS 1858 307AGCUCAUUAAAACAGUCACCAUCAA 31% 15390 CDS 1859 308GCUCAUUAAAACAGUCACCAUCAAA 46% 15391 CDS 1860 309CUCAUUAAAACAGUCACCAUCAAUA 43% 15392 CDS 1861 310UCAUUAAAACAGUCACCAUCAAUGA 58% 15393 CDS 1862 311CAUUAAAACAGUCACCAUCAAUGCA 78% 15394 CDS 1864 312UUAAAACAGUCACCAUCAAUGCAAA 41% 15395 CDS 1865 313UAAAACAGUCACCAUCAAUGCAAGA 80% 15396 CDS 1866 314AAAACAGUCACCAUCAAUGCAAGUA 79% 15397 CDS 1868 315AACAGUCACCAUCAAUGCAAGUUCA 34% 15398 CDS 1912 316AUAUCAAUCCCACAGUACUACUAAA 39% 15399 CDS 1928 317ACUACUAAAAGAACGUUCGACUGAA 39% 15400 CDS/3UTR 1941 318CGUUCGACUGAACUGUAGAAGUCUA 30% 15401 CDS/3UTR 1946 319GACUGAACUGUAGAAGUCUAAUGAA 25% 15402 CDS/3UTR 1949 320UGAACUGUAGAAGUCUAAUGAUCAA 29% 15403 3UTR 2077 321UCCUGUUGCGGAGAAAGGAGUCAUA 45% 15404 3UTR 2082 322UUGCGGAGAAAGGAGUCAUACUU 43% GA 15405 3UTR 2098 323CAUACUUGUGAAGACUUUUAUGU 30% CA 15406 3UTR 2128 324CUAAAGAUUUUGCUGUUGCUGUU 41% AA 15407 3UTR 2141 325UGUUGCUGUUAAGUUUGGAAAAC 29% AA 15408 3UTR 2188 326AGAGAGAAAUGAGUUUUGACGUC 26% UA 15409 3UTR 2235 327UUAUAAGAACGAAAGUAAAGAUGUA 33% 15410 3UTR 2281 328AAGAUGGCAAAAUGCUGAAAGUUUA 28% 15411 3UTR 2305 329UUACACUGUCGAUGUUUCCAAUGCA 46% 15412 3UTR 2446 330GACAUUACCAGUAAUUUCAUGUCUA 24% 15413 3UTR 2581 331CAAAAAGAAGCUGUCUUGGAUUUAA 36% 15414 3UTR 2669 332CUUUUUCACCAAGAGUAUAAACCUA 41% 15415 3UTR 2730 333AUGCCAAAUUUAUUAAGGUGGUG 61% GA 15416 3UTR 2750 334GUGGAGCCACUGCAGUGUUAUCU 39% UA 15417 3UTR 2752 335GGAGCCACUGCAGUGUUAUCUUAAA 45% 15418 3UTR 2802 336CAGAAUUUGUUUAUAUGGCUGGU 49% AA 15419 3UTR 2810 337GUUUAUAUGGCUGGUAACAUGUA 34% AA 15420 3UTR 2963 338UACUCAGAUUUUGCUAUGAGGUU 42% AA 15421 3UTR 2967 339CAGAUUUUGCUAUGAGGUUAAUG 39% AA 15422 3UTR 2970 340AUUUUGCUAUGAGGUUAAUGAAG 43% UA 15423 3UTR 2986 341AAUGAAGUACCAAGCUGUGCUUGAA 40% 15424 3UTR 3064 342AUCACAUUGCAAAAGUAGCAAUGAA 59% 15425 3UTR 3072 343GCAAAAGUAGCAAUGACCUCAUAAA 35% 15426 3UTR 3083 344AAUGACCUCAUAAAAUACCUCUUCA 40% 15427 3UTR 3134 345AAUUUUAUCUCAGUCUUGAAGCCAA 55% 15428 3UTR 3147 346UCUUGAAGCCAAUUCAGUAGGUGCA 52% 15429 3UTR 3157 347AAUUCAGUAGGUGCAUUGGAAUCAA 71% 15430 3UTR 3212 348UUUCUUCUUUUAGCCAUUUUGCU 38% AA 15431 3UTR 3216 349UUCUUUUAGCCAUUUUGCUAAGA 40% GA 15432 3UTR 3225 350CCAUUUUGCUAAGAGACACAGUCUA 36% 15433 3UTR 3278 351UUACUAGUUUUAAGAUCAGAGUU 70% CA 15434 3UTR 3313 352ACUCUGCCUAUAUUUUCUUACCUGA 56% 15435 3UTR 3335 353UGAACUUUUGCAAGUUUUCAGGU 64% AA 15436 3UTR 3336 354GAACUUUUGCAAGUUUUCAGGUA 62% AA 15437 3UTR 3351 355UUCAGGUAAACCUCAGCUCAGGACA 62% 15438 3UTR 3360 356ACCUCAGCUCAGGACUGCUAUUUAA 53% 15439 3UTR 3441 357CUUAUUUUAAGUGAAAAGCAGAGAA 83% 15440 3UTR 3489 358UAUCUGUAACCAAGAUGGAUGCAAA 93% 15441 3UTR 3662 359UUUUCCACAUCUCAUUGUCACUGAA 36% 15442 3UTR 3668 360ACAUCUCAUUGUCACUGACAUUUAA 40% 15443 3UTR 3735 361GUCUUAUUAGGACACUAUGGUUA 40% UA 15444 3UTR 3737 362CUUAUUAGGACACUAUGGUUAUA 37% AA 15445 3UTR 3738 363UUAUUAGGACACUAUGGUUAUAA 38% AA 15446 3UTR 3752 364UGGUUAUAAACUGUGUUUAAGCC 28% UA 15447 3UTR 3919 365AUAUUUAAGGUUGAAUGUUUGUC 40% CA 15448 3UTR 3961 366CUAGCCCACAAAGAAUAUUGUCUCA 47% 15449 3UTR 3981 367UCUCAUUAGCCUGAAUGUGCCAUAA 56% 15450 3UTR 3994 368AAUGUGCCAUAAGACUGACCUUUUA 52%

TABLE 3 PTGS2 sd-rxRNA Duplex ID ID Sequence SEQ ID NO 17388 17062G.A.A.A.A.mC.mU.G.mC.mU.mC.A.A.Chl 369 17063P.mU.fU.G.A.G.fC.A.G.fU.fU.fU.fU.fC*fU*fC*fC*A*fU*A 370 17389 17064A.mC.mC.mU.mC.mU.mC.mC.mU.A.mU.mU.A.Chl 371 17065P.mU.A.A.fU.A.G.G.A.G.A.G.G.fU*fU*A*G*A*G*A 372 17390 17066mU.mC.mC.A.mC.mC.A.A.mC.mU.mU.A.A.Chl 373 17068P.mU.fU.A.A.G.fU.fU.G.G.fU.G.G.A*fC*fU*G*fU*fC*A 374 17391 17067G.mU.mC.mC.A.mC.mC.A.A.mC.mU.mU.A.A.Chl 375 17068P.mU.fU.A.A.G.fU.fU.G.G.fU.G.G.A*fC*fU*G*fU*fC*A 376 17392 17069mC.mU.mC.mC.mU.A.mU.mU.A.mU.A.mC.A.Chl 377 17070P.mU.G.fU.A.fU.A.A.fU.A.G.G.A.G*A*G*G*fU*fU*A 378 17393 17071G.A.mU.mC.A.mC.A.mU.mU.mU.G.A.A.Chl 379 17073P.mU.fU.fC.A.A.A.fU.G.fU.G.A.fU.fC*fU*G*G*A*fU*G 380 17394 17072A.G.A.mU.mC.A.mC.A.mU.mU.mU.G.A.A.Chl 381 17073P.mU.fU.fC.A.A.A.fU.G.fU.G.A.fU.fC*fU*G*G*A*fU*G 382 17395 17074A.A.mC.mC.mU.mC.mU.mC.mC.mU.A.mU.A.Chl 383 17075P.mU.A.fU.A.G.G.A.G.A.G.G.fU.fU*A*G*A*G*A*A 384 17396 17076G.mU.mU.G.A.mC.A.mU.mC.mC.A.G.A.Chl 385 17077P.mU.fC.fU.G.G.A.fU.G.fU.fC.A.A.fC*A*fC*A*fU*A*A 386 17397 17078mC.mC.mU.mU.mC.mC.mU.mU.mC.G.A.A.A.Chl 387 17079P.mU.fU.fU.fC.G.A.A.G.G.A.A.G.G*G*A*A*fU*G*U 388 17398 17080A.mC.mU.mC.mC.A.A.A.mC.A.mC.A.A.Chl 389 17082P.mU.fU.G.fU.G.fU.fU.fU.G.G.A.G.fU*G*G*G*fU*fU*U 390 17399 17081mC.A.mC.mU.mC.mC.A.A.A.mC.A.mC.A.A.Chl 391 17082P.mU.fU.G.fU.G.fU.fU.fU.G.G.A.G.fU*G*G*G*fU*fU*U 392 17400 17083mC.A.mC.mU.mC.mC.A.A.A.mC.A.mC.A.Chl 393 17084P.mUGfUGfUfUfUGGAGfUG*G*G*fU*fU*fU*C 394 17401 17085mC.mC.A.mC.mC.A.A.mC.mU.mU.A.mCA.Chl 395 17087P.mUGfUAAGfUfUGGfUGG*A*fC*fU*G*fU*C 396 17402 17086mU.mC.mC.A.mC.mC.A.A.mC.mU.mU.A.mCA.Chl 397 17087P.mUGfUAAGfUfUGGfUGG*A*fC*fU*G*fU*C 398 17403 17088A.A.mU.A.mC.mC.A.G.mU.mC.mU.mU.A.Chl 399 17089P.mU.A.A.G.A.fC.fU.G.G.fU.A.fU.fU*fU*fC*A*fU*fC*U 400 17404 17090G.A.mC.mC.A.G.mU.A.mU.A.A.G.A.Chl 401 17091P.mU.fC.fU.fU.A.fU.A.fC.fU.G.G.fU.fC*A*A*A*fU*fC*C 402 17405 17092G.mU.mC.mU.mU.mU.mU.A.A.mU.G.A.A.Chl 403 17093P.mU.fU.fC.A.fU.fU.A.A.A.A.G.A.fC*fU*G*G*fU*A*U 404 17406 17094A.A.mU.mU.mU.mC.A.mU.G.mU.mC.mU.A.Chl 405 17095P.mU.A.G.A.fC.A.fU.G.A.A.A.fU.fU*A*fC*fU*G*G*U 406 17407 17096A.mU.mC.A.mC.A.mU.mU.mU.G.A.mU.A.Chl 407 17098P.mU.A.fU.fC.A.A.A.fU.G.fU.G.A.fU*fC*fU*G*G*A*U 408 17408 17097G.A.mU.mC.A.mC.A.mU.mU.mU.G.A.mU.A.Chl 409 17098P.mU.A.fU.fC.A.A.A.fU.G.fU.G.A.fU*fC*fU*G*G*A*U 410 17409 17099mU.mC.mC.A.G.A.mU.mC.A.mC.A.mU.A.Chl 411 17100P.mU.A.fU.G.fU.G.A.fU.fC.fU.G.G.A*fU*G*fU*fC*A*A 412 17410 17101mU.A.mC.mU.G.A.mU.A.G.G.A.G.A.Chl 413 17102P.mU.fC.fU.fC.fC.fU.A.fU.fC.A.G.fU.A*fU*fU*A*G*fC*C 414 17411 17103G.mU.G.mC.A.A.mC.A.mC.mU.fU.G.A.Chl 415 17104P.mU.fC.A.A.G.fU.G.fU.fU.G.mC.A.fC*A*fU*A*A*fU*C 416 17412 17105A.mC.mC.A.G.mU.A.mU.A.A.G.mU.A.Chl 417 17106P.mU.A.fC.fU.fU.A.fU.A.fC.fU.G.G.fU*fC*A*A*A*fU*C 418 17413 17107G.A.A.G.mU.mC.mU.A.A.mU.G.A.A.Chl 419 17108P.mU.fU.fC.A.fU.fU.A.G.A.fC.mU.fU.fC*fU*A*fC*A*G*U 420 17414 17109A.A.G.A.A.G.A.A.A.G.mU.mU.A.Chl 421 17110P.mU.A.A.fC.fU.fU.fU.fC.fU.fU.fC.fU.fU*A*G*A*A*G*C 422 17415 17111mU.mC.A.mC.A.mU.mU.mU.G.AmU.mU.A.Chl 423 17113P.mU.A.A.fU.fC.A.A.A.fU.G.fUG.A*fU*fC*fU*G*G*A 424 17416 17112A.mU.mC.A.mC.A.mU.mU.mU.G.AmU.mU.A.Chl 425 17113P.mU.A.A.fU.fC.A.A.A.fU.G.fUG.A*fU*fC*fU*G*G*A 426 17417 17114A.mC.A.mU.mU.mU.G.A.mU.mUG.A.A.Chl 427 17116P.mU.fU.fC.A.A.fU.fC.A.A.A.fUG.fU*G*A*fU*fC*fU*G 428 17418 17115mC.A.mC.A.mU.mU.mU.G.A.mU.mUG.A.A.Chl 429 17116P.mU.fU.fC.A.A.fU.fC.A.A.A.fUG.fU*G*A*fU*fC*fU*G 430 17419 17117A.mU.mU.mU.G.A.mU.mU.G.AmC.A.A.Chl 431 17119P.mU.fU.G.fU.fC.A.A.fU.fC.A.AA.fU*G*fU*G*A*fU*C 432 17420 17118mC.A.mU.mU.mU.G.A.mU.mU.G.AmC.A.A.Chl 433 17119P.mU.fU.G.fU.fC.A.A.fU.fC.A.AA.fU*G*fU*G*A*fU*C 434 17421 17120mC.A.mU.mC.mU.G.mC.A.A.mU.A.A.A.Chl 435 17122P.mU.fU.fU.A.fU.fU.G.fC.A.G.A.fU.G*A*G*A*G*A*C 436 17422 17121mU.mC.A.mU.mC.mU.G.mC.A.A.mU.A.A.A.Chl 437 17122P.mU.fU.fU.A.fU.fU.G.fC.A.G.A.fU.G*A*G*A*G*A*C 438

TABLE 4 TGFB1 sd-rxRNA Target Gene hTGFB1 Duplex ID Single Strand IDsd-rxRNA sequence SEQ ID NO 18454 17491mC.A.mC.A.G.mC.A.mU.A.mU.A.mU.A.Chl 439 17492P.mU.A.fU.A.fU.A.fU.G.fC.fU.G.fU.G*fU*G*fU*A*fC*U 440 18455 17493mC.A.G.mC.A.mU.A.mU.A.mU.A.mU.A.Chl 441 17494P.mU.A.fU.A.fU.A.fU.A.fU.G.fC.fU.G*fU*G*fU*G*fU*A 442 18456 17495G.mU.A.mC.A.mU.mU.G.A.mC.mU.mU.A.Chl 443 17497P.mU.A.A.G.fU.fC.A.A.fU.G.fU.A.fC*A*G*fC*fU*G*C 444 18457 17496mU.G.mU.A.mC.A.mU.mU.G.A.mC.mU.mU.A.Chl 445 17497P.mU.A.A.G.fU.fC.A.A.fU.G.fU.A.fC*A*G*fC*fU*G*C 446 18458 17498A.A.mC.mU.A.mU.mU.G.mC.mU.mU.mC.A.Chl 447 17500P.mU.G.A.A.G.fC.A.A.fU.A.G.fU.fU*G*G*fU*G*fU*C 448 18459 17499mC.A.A.mC.mU.A.mU.mU.G.mC.mU.mU.mC.A.Chl 449 17500P.mU.G.A.A.G.fC.A.A.fU.A.G.fU.fU*G*G*fU*G*fU*C 450 18460 17501G.mC.A.mU.A.mU.A.mU.A.mU.G.mU.A.Chl 451 17502P.mU.A.fC.A.fU.A.fU.A.fU.A.fU.G.fC*fU*G*fU*G*fU*G 452 18461 17503mU.G.mU.A.mC.A.mU.mU.G.A.mC.mU.A.Chl 453 17505P.mU.A.G.fU.fC.A.A.fU.G.fU.A.fC.A*G*fC*fU*G*fC*C 454 18462 17504mC.mU.G.mU.A.mC.A.mU.mU.G.A.mC.mU.A.Chl 455 17505P.mU.A.G.fU.fC.A.A.fU.G.fU.A.fC.A*G*fC*fU*G*fC*C 456 18463 17506A.G.mC.A.mU.A.mU.A.mU.A.mU.G.A.Chl 457 17507P.mU.fC.A.fU.A.fU.A.fU.A.fU.G.fC.fU*G*fU*G*fU*G*U 458 18464 17508mC.A.G.mC.A.A.mC.A.A.mU.mU.mC.A.Chl 459 17509P.mU.G.A.A.fU.fU.G.fU.fU.G.fC.fU.G*fU*A*fU*fU*fU*C 460 18465 17510mC.A.mU.A.mU.A.mU.A.mU.G.mU.mU.A.Chl 461 17511P.mU.A.A.fC.A.fU.A.fU.A.fU.A.fU.G*fC*fU*G*fU*G*U 462 18466 17512mU.mU.G.mC.mU.mU.mC.A.G.mC.mU.mC.A.Chl 463 17514P.mU.G.A.G.fC.fU.G.A.A.G.fC.A.A*fU*A*G*fU*fU*G 464 18467 17513A.mU.mU.G.mC.mU.mU.mC.A.G.mC.mU.mC.A.Chl 465 17514P.mU.G.A.G.fC.fU.G.A.A.G.fC.A.A*fU*A*G*fU*fU*G 466 18468 17515A.mC.A.G.mC.A.mU.A.mU.A.mU.A.A.Chl 467 17516P.mU.fU.A.fU.A.fU.A.fU.G.fC.fU.G.fU*G*fU*G*fU*A*C 468 18469 17517A.mU.mU.G.mC.mU.mU.mC.A.G.mC.mU.A.Chl 469 17519P.mU.A.G.fC.fU.G.A.A.G.fC.A.A.fU*A*G*fU*fU*G*G 470 18470 17518mU.A.mU.mU.G.mC.mU.mU.mC.A.G.mC.mU.A.Chl 471 17519P.mU.A.G.fC.fU.G.A.A.G.fC.A.A.fU*A*G*fU*fU*G*G 472 18471 17520mC.A.G.A.G.mU.A.mC.A.mC.A.mC.A.Chl 473 17521P.mU.G.fU.G.fU.G.fU.A.fC.fU.fC.fU.G*C*fU*fU*G*A*A 474 18472 17522mU.mC.A.A.G.mC.A.G.A.G.mU.A.A.Chl 475 17523P.mU.fU.A.fC.fU.fC.fU.G.fC.fU.fU.G.A*A*fC*fU*fU*G*U 476 18473 17524A.G.mC.A.G.A.G.mU.A.mC.A.mC.A.Chl 477 17525P.mU.G.fU.G.fU.A.fC.fU.fC.fU.G.fC.fU*fU*G*A*A*fC*U 478 18474 17526G.A.mC.A.A.G.mU.mU.mC.A.A.G.A.Chl 479 17527P.mU.fC.fU.fU.G.A.A.fC.fU.fU.G.fU.fC*A*fU*A*G*A*U 480 18475 17528mC.mU.A.mU.G.A.mC.A.A.G.mU.mU.A.Chl 481 17529P.mU.A.A.fC.fU.fU.G.fU.fC.A.fU.A.G*A*fU*fU*fU*fC*G 482 18476 17530G.mC.A.G.A.G.mU.A.mC.A.mC.A.A.Chl 483 17531P.mU.fU.G.fU.G.fU.A.fC.fU.fC.fU.G.fC*fU*fU*G*A*A*C 484 18477 17532mU.G.A.mC.A.A.G.mU.mU.mCA.A.A.Chl 485 17533P.mU.fU.fU.G.A.A.fC.fU.fU.GfU.fC.A*fU*A*G*A*fU*U 486 18478 17534mU.A.mC.A.mC.A.mC.A.G.mCA.mU.A.Chl 487 17535P.mU.A.fU.G.fC.fU.G.fU.G.fUG.fU.A*fC*fU*fC*fU*G*C 488 18479 17536A.A.mC.G.A.A.A.mU.mC.mUA.mU.A.Chl 489 17537P.mU.A.fU.A.G.A.fU.fU.fU.fCG.fU.fU*G*fU*G*G*G*U 490 18480 17538mU.mU.G.A.mC.mU.mU.mC.mC.GmC.A.A.Chl 491 17539P.mU.fU.G.fC.G.G.A.A.G.fUfC.A.A*fU*G*fU*A*fC*A 492 18481 17540A.mC.A.A.mC.G.A.A.A.mUmC.mU.A.Chl 493 17541P.mU.A.G.A.fU.fU.fU.fC.G.fUfU.G.fU*G*G*G*fU*fU*U 494 18482 17542mU.mC.A.A.mC.A.mC.A.mU.mCA.G.A.Chl 495 17543P.mU.fC.fU.G.A.fU.G.fU.G.fUfU.G.A*A*G*A*A*fC*A 496 18483 17544A.mC.A.A.G.mU.mU.mC.A.AG.mC.A.Chl 497 17545P.mU.G.fC.fU.fU.G.A.A.fC.fUfU.G.fU*fC*A*fU*A*G*A 498 18484 17546A.mU.mC.mU.A.mU.G.A.mC.AA.G.A.Chl 499 17547P.mU.fC.fU.fU.G.fU.fC.A.fU.AG.A.fU*fU*fU*fC*G*fU*U 500 Rat TargetingTGFB1 18715 18691 G.A.A.A.mU.A.mU.A.G.mC.A.A.A-chol 503 18692P.mU.fU.fU.G.fC.fU.A.fU.A.fU.fU.fU.fC*fU*G*G*fU*A*G 504 18716 18693G.A.A.mC.mU.mC.mU.A.mC.mC.A.G.A-chol 505 18694P.mU.fC.fU.G.G.fU.A.G.A.G.fU.fU.fC*fU*A*fC*G*fU*G 506 18717 18695G.mC.A.A.A.G.A.mU.A.A.mU.G.A-chol 507 18696P.mU.fC.A.fU.fU.A.fU.fC.fU.fU.fU.G.fC*fU*G*fU*fC*A*C 508 18718 18697A.A.mC.mU.mC.mU.A.mC.mC.A.G.A.A-chol 509 18698P.mU.fU.fC.fU.G.G.fU.A.G.A.G.fU.fU*fC*fU*A*fC*G*U 510 18719 18699A.mC.mU.mC.mU.A.mC.mC.A.G.A.A.A-chol 511 18700P.mU.fU.fU.fC.fU.G.G.fU.A.G.A.G.fU*fU*fC*fU*A*fC*G 512 18720 18701A.mC.A.G.mC.A.A.A.G.A.mU.A.A-chol 513 18702P.mU.fU.A.fU.fC.fU.fU.fU.G.fC.fU.G.fU*fC*A*fC*A*A*G 514 18721 18703mC.A.A.mU.mC.mU.A.mU.G.A.mC.A.A-chol 515 18704P.mU.fU.G.fU.fC.A.fU.A.G.A.fU.fU.G*fC*G*fU*fU*G*U 516 18722 18705A.G.A.mU.mU.mC.A.A.G.mU.mC.A.A-chol 517 18706P.mU.fU.G.A.fC.fU.fU.G.A.A.fU.fC.fU*fC*fU*G*fC*A*G 518 18723 18707mC.mU.G.mU.G.G.A.G.mC.A.A.mC.A-chol 519 18708P.mU.G.fU.fU.G.fC.fU.fC.fC.A.fC.A.G*fU*fU*G*A*fC*U 520 18724 18709mU.G.A.mC.A.G.mC.A.A.A.G.A.A-chol 521 18710P.mU.fU.fC.fU.fU.fU.G.fC.fU.G.fU.fC.A*fC*A*A*G*A*G 522 18725 18711A.mU.G.A.mC.A.A.A.A.mC.mC.A.A-chol 523 18712P.mU.fU.G.G.fU.fU.fU.fU.G.fU.fC.A.fU*A*G*A*fU*fU*G 524 18726 18713G.A.G.A.mU.mU.mC.A.A.G.mU.mC.A-chol 525 18714P.mU.G.A.fC.fU.fU.G.A.A.fU.fC.fU.fC*fU*G*fC*A*G*G 526

TABLE 5 Inhibition of gene expression with hTGFB1 ori sequences %Expression Target Gene Gene SEQ ID hTGFB1 0.025 nM HeLa Duplex ID RegionRef Pos NO Sense Sequence cells 15732 CDS 954 527CGCGGGACUAUCCACCUGCAAGACA 57.3% 15733 CDS 956 528CGGGACUAUCCACCUGCAAGACUAA 38.2% 15734 CDS 957 529GGGACUAUCCACCUGCAAGACUAUA 49.1% 15735 CDS 961 530CUAUCCACCUGCAAGACUAUCGACA 34.9% 15736 CDS 962 531UAUCCACCUGCAAGACUAUCGACAA 39.4% 15737 CDS 964 532UCCACCUGCAAGACUAUCGACAUGA 44.4% 15738 CDS 965 533CCACCUGCAAGACUAUCGACAUGGA 53.3% 15739 CDS 966 534CACCUGCAAGACUAUCGACAUGGAA 52.8% 15740 CDS 967 535ACCUGCAAGACUAUCGACAUGGAGA 46.2% 15741 CDS 968 536CCUGCAAGACUAUCGACAUGGAGCA 48.1% 15742 CDS 1209 537AAUGGUGGAAACCCACAACGAAAUA 36.7% 15743 CDS 1210 538AUGGUGGAAACCCACAACGAAAUCA 28.8% 15744 CDS 1211 539UGGUGGAAACCCACAACGAAAUCUA 23.1% 15745 CDS 1212 540GGUGGAAACCCACAACGAAAUCUAA 13.2% 15746 CDS 1213 541GUGGAAACCCACAACGAAAUCUAUA 21.1% 15747 CDS 1214 542UGGAAACCCACAACGAAAUCUAUGA 28.7% 15748 CDS 1215 543GGAAACCCACAACGAAAUCUAUGAA 32.9% 15749 CDS 1216 544GAAACCCACAACGAAAUCUAUGACA 41.5% 15750 CDS 1217 545AAACCCACAACGAAAUCUAUGACAA 29.9% 15751 CDS 1218 546AACCCACAACGAAAUCUAUGACAAA 16.4% 15752 CDS 1219 547ACCCACAACGAAAUCUAUGACAAGA 23.3% 15753 CDS 1220 548CCCACAACGAAAUCUAUGACAAGUA 37.5% 15754 CDS 1221 549CCACAACGAAAUCUAUGACAAGUUA 19.1% 15755 CDS 1222 550CACAACGAAAUCUAUGACAAGUUCA 14.4% 15756 CDS 1224 551CAACGAAAUCUAUGACAAGUUCAAA 20.1% 15757 CDS 1225 552AACGAAAUCUAUGACAAGUUCAAGA 18.3% 15758 CDS 1226 553ACGAAAUCUAUGACAAGUUCAAGCA 23.2% 15759 CDS 1227 554CGAAAUCUAUGACAAGUUCAAGCAA 29.0% 15760 CDS 1228 555GAAAUCUAUGACAAGUUCAAGCAGA 15.6% 15761 CDS 1229 556AAAUCUAUGACAAGUUCAAGCAGAA 32.3% 15762 CDS 1230 557AAUCUAUGACAAGUUCAAGCAGAGA 36.1% 15763 CDS 1231 558AUCUAUGACAAGUUCAAGCAGAGUA 30.6% 15764 CDS 1232 559UCUAUGACAAGUUCAAGCAGAGUAA 24.9% 15765 CDS 1233 560CUAUGACAAGUUCAAGCAGAGUACA 15.9% 15766 CDS 1234 561UAUGACAAGUUCAAGCAGAGUACAA 31.2% 15767 CDS 1235 562AUGACAAGUUCAAGCAGAGUACACA 17.2% 15768 CDS 1236 563UGACAAGUUCAAGCAGAGUACACAA 23.5% 15769 CDS 1237 564GACAAGUUCAAGCAGAGUACACACA 24.5% 15770 CDS 1238 565ACAAGUUCAAGCAGAGUACACACAA 38.5% 15771 CDS 1240 566AAGUUCAAGCAGAGUACACACAGCA 38.7% 15772 CDS 1241 567AGUUCAAGCAGAGUACACACAGCAA 34.3% 15773 CDS 1242 568GUUCAAGCAGAGUACACACAGCAUA 20.8% 15774 CDS 1243 569UUCAAGCAGAGUACACACAGCAUAA 33.4% 15775 CDS 1244 570UCAAGCAGAGUACACACAGCAUAUA 19.6% 15776 CDS 1245 571CAAGCAGAGUACACACAGCAUAUAA 25.5% 15777 CDS 1246 572AAGCAGAGUACACACAGCAUAUAUA 12.8% 15778 CDS 1247 573AGCAGAGUACACACAGCAUAUAUAA 27.6% 15779 CDS 1248 574GCAGAGUACACACAGCAUAUAUAUA 15.9% 15780 CDS 1249 575CAGAGUACACACAGCAUAUAUAUGA 24.1% 15781 CDS 1250 576AGAGUACACACAGCAUAUAUAUGUA 22.6% 15782 CDS 1251 577GAGUACACACAGCAUAUAUAUGUUA 26.7% 15783 CDS 1252 578AGUACACACAGCAUAUAUAUGUUCA 66.6% 15784 CDS 1254 579UACACACAGCAUAUAUAUGUUCUUA 33.6% 15785 CDS 1262 580GCAUAUAUAUGUUCUUCAACACAUA 40.4% 15786 CDS 1263 581CAUAUAUAUGUUCUUCAACACAUCA 42.5% 15787 CDS 1264 582AUAUAUAUGUUCUUCAACACAUCAA 27.2% 15788 CDS 1265 583UAUAUAUGUUCUUCAACACAUCAGA 23.2% 15789 CDS 1266 584AUAUAUGUUCUUCAACACAUCAGAA 35.5% 15790 CDS 1267 585UAUAUGUUCUUCAACACAUCAGAGA 34.6% 15791 CDS 1268 586AUAUGUUCUUCAACACAUCAGAGCA 29.7% 15792 CDS 1269 587UAUGUUCUUCAACACAUCAGAGCUA 35.4% 15793 CDS 1270 588AUGUUCUUCAACACAUCAGAGCUCA 35.2% 15794 CDS 1335 589GCUGCGUCUGCUGAGGCUCAAGUUA 28.0% 15795 CDS 1336 590CUGCGUCUGCUGAGGCUCAAGUUAA 32.1% 15796 CDS 1337 591UGCGUCUGCUGAGGCUCAAGUUAAA 25.5% 15797 CDS 1338 592GCGUCUGCUGAGGCUCAAGUUAAAA 59.7% 15798 CDS 1339 593CGUCUGCUGAGGCUCAAGUUAAAAA 52.8% 15799 CDS 1340 594GUCUGCUGAGGCUCAAGUUAAAAGA 47.9% 15800 CDS 1341 595UCUGCUGAGGCUCAAGUUAAAAGUA 49.8% 15801 CDS 1342 596CUGCUGAGGCUCAAGUUAAAAGUGA 50.7% 15802 CDS 1343 597UGCUGAGGCUCAAGUUAAAAGUGGA 43.4% 15803 CDS 1344 598GCUGAGGCUCAAGUUAAAAGUGGAA 52.6% 15804 CDS 1345 599CUGAGGCUCAAGUUAAAAGUGGAGA 73.3% 15805 CDS 1346 600UGAGGCUCAAGUUAAAAGUGGAGCA 58.0% 15806 CDS 1347 601GAGGCUCAAGUUAAAAGUGGAGCAA 64.9% 15807 CDS 1348 602AGGCUCAAGUUAAAAGUGGAGCAGA 68.1% 15808 CDS 1349 603GGCUCAAGUUAAAAGUGGAGCAGCA 73.8% 15809 CDS 1350 604GCUCAAGUUAAAAGUGGAGCAGCAA 78.8% 15810 CDS 1351 605CUCAAGUUAAAAGUGGAGCAGCACA 76.6% 15811 CDS 1352 606UCAAGUUAAAAGUGGAGCAGCACGA 72.9% 15812 CDS 1369 607CAGCACGUGGAGCUGUACCAGAAAA 69.8% 15813 CDS 1370 608AGCACGUGGAGCUGUACCAGAAAUA 69.7% 15814 CDS 1371 609GCACGUGGAGCUGUACCAGAAAUAA 73.3% 15815 CDS 1372 610CACGUGGAGCUGUACCAGAAAUACA 55.0% 15816 CDS 1373 611ACGUGGAGCUGUACCAGAAAUACAA 63.8% 15817 CDS 1374 612CGUGGAGCUGUACCAGAAAUACAGA 85.7% 15818 CDS 1375 613GUGGAGCUGUACCAGAAAUACAGCA 85.0% 15819 CDS 1376 614UGGAGCUGUACCAGAAAUACAGCAA 82.5% 15820 CDS 1377 615GGAGCUGUACCAGAAAUACAGCAAA 43.1% 15821 CDS 1378 616GAGCUGUACCAGAAAUACAGCAACA 58.5% 15822 CDS 1379 617AGCUGUACCAGAAAUACAGCAACAA 48.1% 15823 CDS 1380 618GCUGUACCAGAAAUACAGCAACAAA 48.1% 15824 CDS 1381 619CUGUACCAGAAAUACAGCAACAAUA 35.0% 15825 CDS 1382 620UGUACCAGAAAUACAGCAACAAUUA 36.4% 15826 CDS 1383 621GUACCAGAAAUACAGCAACAAUUCA 24.6% 15827 CDS 1384 622UACCAGAAAUACAGCAACAAUUCCA 33.4% 15828 CDS 1385 623ACCAGAAAUACAGCAACAAUUCCUA 121.5% 15829 CDS 1386 624CCAGAAAUACAGCAACAAUUCCUGA 62.1% 15830 CDS 1387 625CAGAAAUACAGCAACAAUUCCUGGA 98.3% 15831 CDS 1390 626AAAUACAGCAACAAUUCCUGGCGAA 36.6% 15832 CDS 1391 627AAUACAGCAACAAUUCCUGGCGAUA 39.5% 15833 CDS 1392 628AUACAGCAACAAUUCCUGGCGAUAA 40.0% 15834 CDS 1393 629UACAGCAACAAUUCCUGGCGAUACA 89.4% 15835 CDS 1394 630ACAGCAACAAUUCCUGGCGAUACCA 62.3% 15836 CDS 1396 631AGCAACAAUUCCUGGCGAUACCUCA 41.0% 15837 CDS 1441 632AGCGACUCGCCAGAGUGGUUAUCUA 31.2% 15838 CDS 1442 633GCGACUCGCCAGAGUGGUUAUCUUA 46.2% 15839 CDS 1443 634CGACUCGCCAGAGUGGUUAUCUUUA 46.8% 15840 CDS 1444 635GACUCGCCAGAGUGGUUAUCUUUUA 50.6% 15841 CDS 1445 636ACUCGCCAGAGUGGUUAUCUUUUGA 50.8% 15842 CDS 1446 637CUCGCCAGAGUGGUUAUCUUUUGAA 71.8% 15843 CDS 1447 638UCGCCAGAGUGGUUAUCUUUUGAUA 43.7% 15844 CDS 1448 639CGCCAGAGUGGUUAUCUUUUGAUGA 42.1% 15845 CDS 1449 640GCCAGAGUGGUUAUCUUUUGAUGUA 31.0% 15846 CDS 1450 641CCAGAGUGGUUAUCUUUUGAUGUCA 46.0% 15847 CDS 1451 642CAGAGUGGUUAUCUUUUGAUGUCAA 40.2% 15848 CDS 1452 643AGAGUGGUUAUCUUUUGAUGUCACA 38.5% 15849 CDS 1453 644GAGUGGUUAUCUUUUGAUGUCACCA 67.4% 15850 CDS 1454 645AGUGGUUAUCUUUUGAUGUCACCGA 57.4% 15851 CDS 1455 646GUGGUUAUCUUUUGAUGUCACCGGA 40.6% 15852 CDS 1456 647UGGUUAUCUUUUGAUGUCACCGGAA 70.5% 15853 CDS 1457 648GGUUAUCUUUUGAUGUCACCGGAGA 82.8% 15854 CDS 1458 649GUUAUCUUUUGAUGUCACCGGAGUA 74.8% 15855 CDS 1459 650UUAUCUUUUGAUGUCACCGGAGUUA 86.8% 15856 CDS 1460 651UAUCUUUUGAUGUCACCGGAGUUGA 76.5% 15857 CDS 1551 652CAGCAGGGAUAACACACUGCAAGUA 70.5% 15858 CDS 1552 653AGCAGGGAUAACACACUGCAAGUGA 60.5% 15859 CDS 1553 654GCAGGGAUAACACACUGCAAGUGGA 43.5% 15860 CDS 1554 655CAGGGAUAACACACUGCAAGUGGAA 56.3% 15861 CDS 1555 656AGGGAUAACACACUGCAAGUGGACA 63.9% 15862 CDS 1556 657GGGAUAACACACUGCAAGUGGACAA 66.9% 15863 CDS 1558 658GAUAACACACUGCAAGUGGACAUCA 62.2% 15864 CDS 1559 659AUAACACACUGCAAGUGGACAUCAA 40.5% 15865 CDS 1560 660UAACACACUGCAAGUGGACAUCAAA 57.9% 15866 CDS 1610 661ACCUGGCCACCAUUCAUGGCAUGAA 69.4% 15867 CDS 1611 662CCUGGCCACCAUUCAUGGCAUGAAA 49.1% 15868 CDS 1612 663CUGGCCACCAUUCAUGGCAUGAACA 31.9% 15869 CDS 1705 664CGAGCCCUGGACACCAACUAUUGCA 56.4% 15870 CDS 1706 665GAGCCCUGGACACCAACUAUUGCUA 42.6% 15871 CDS 1707 666AGCCCUGGACACCAACUAUUGCUUA 29.8% 15872 CDS 1708 667GCCCUGGACACCAACUAUUGCUUCA 19.8% 15873 CDS 1709 668CCCUGGACACCAACUAUUGCUUCAA 37.7% 15874 CDS 1710 669CCUGGACACCAACUAUUGCUUCAGA 44.0% 15875 CDS 1711 670CUGGACACCAACUAUUGCUUCAGCA 35.8% 15876 CDS 1712 671UGGACACCAACUAUUGCUUCAGCUA 31.5% 15877 CDS 1713 672GGACACCAACUAUUGCUUCAGCUCA 27.3% 15878 CDS 1714 673GACACCAACUAUUGCUUCAGCUCCA 44.7% 15879 CDS 1715 674ACACCAACUAUUGCUUCAGCUCCAA 44.9% 15880 CDS 1754 675GCGUGCGGCAGCUGUACAUUGACUA 23.9% 15881 CDS 1755 676CGUGCGGCAGCUGUACAUUGACUUA 18.3% 15882 CDS 1756 677GUGCGGCAGCUGUACAUUGACUUCA 41.2% 15883 CDS 1757 678UGCGGCAGCUGUACAUUGACUUCCA 26.4% 15884 CDS 1759 679CGGCAGCUGUACAUUGACUUCCGCA 28.0% 15885 CDS 1760 680GGCAGCUGUACAUUGACUUCCGCAA 22.8% 15886 CDS 1761 681GCAGCUGUACAUUGACUUCCGCAAA 34.1% 15887 CDS 1762 682CAGCUGUACAUUGACUUCCGCAAGA 36.3% 15888 CDS 1763 683AGCUGUACAUUGACUUCCGCAAGGA 84.1% 15889 CDS 1849 684UGCCCCUACAUUUGGAGCCUGGACA 93.0% 15890 CDS 1889 685UCCUGGCCCUGUACAACCAGCAUAA 51.7% 15891 CDS 1890 686CCUGGCCCUGUACAACCAGCAUAAA 71.9% 15892 CDS 1891 687CUGGCCCUGUACAACCAGCAUAACA 36.1% 15893 CDS 1997 688AGGUGGAGCAGCUGUCCAACAUGAA 60.9% 15894 3UTR 2115 689CAUGGGGGCUGUAUUUAAGGACACA 57.2% 15895 3UTR 2155 690CCUGGGGCCCCAUUAAAGAUGGAGA 86.0% 15896 3UTR 2156 691CUGGGGCCCCAUUAAAGAUGGAGAA 73.3% 15897 3UTR 2157 692UGGGGCCCCAUUAAAGAUGGAGAGA 68.8% 15898 3UTR 2158 693GGGGCCCCAUUAAAGAUGGAGAGAA 65.8% 15899 3UTR 2159 694GGGCCCCAUUAAAGAUGGAGAGAGA 42.7% 15900 3UTR 2160 695GGCCCCAUUAAAGAUGGAGAGAGGA 34.4% 15901 3UTR 2161 696GCCCCAUUAAAGAUGGAGAGAGGAA 56.0% 15902 3UTR 2162 697CCCCAUUAAAGAUGGAGAGAGGACA 74.9% 15903 3UTR 2163 698CCCAUUAAAGAUGGAGAGAGGACUA 79.6% 15904 3UTR 2180 699GAGGACUGCGGAUCUCUGUGUCAUA 98.3% 15905 3UTR 2275 700CUCCUGCCUGUCUGCACUAUUCCUA 100.2% 15906 3UTR 2276 701UCCUGCCUGUCUGCACUAUUCCUUA 103.8% 15907 3UTR 2277 702CCUGCCUGUCUGCACUAUUCCUUUA 110.4% 15908 3UTR 2278 703CUGCCUGUCUGCACUAUUCCUUUGA 105.2% 15909 3UTR 2279 704UGCCUGUCUGCACUAUUCCUUUGCA 118.8% 15910 3UTR 2325 705CAGUGGGGAACACUACUGUAGUUAA 112.2% 15911 3UTR 2326 706AGUGGGGAACACUACUGUAGUUAGA 107.7% 15912 3UTR 2327 707GUGGGGAACACUACUGUAGUUAGAA 108.6% 15913 3UTR 2328 708UGGGGAACACUACUGUAGUUAGAUA N/A

TABLE 6 Inhibition of gene expression with hTGFB2 ori sequences OligoGene % Expression 25-mer Sense Strand (position id Region Ref Pos A5490.1 nM 25 of SS, replaced with A) SEQ ID NO 15451 5UTR/CDS 651 98%UUUUAAAAAAUGCACUACUGUGUGC 709 15452 CDS 654 102.2%UAAAAAAUGCACUACUGUGUGCUGA 710 15453 CDS 730 83.7%GCAGCACACUCGAUAUGGACCAGUU 711 15454 CDS 732 80.3%AGCACACUCGAUAUGGACCAGUUCA 712 15455 CDS 733 79.6%GCACACUCGAUAUGGACCAGUUCAU 713 15456 CDS 734 89.1%CACACUCGAUAUGGACCAGUUCAUG 714 15457 CDS 735 87.8%ACACUCGAUAUGGACCAGUUCAUGC 715 15458 CDS 736 95.3%CACUCGAUAUGGACCAGUUCAUGCG 716 15459 CDS 847 103.8%UCCCCCCGGAGGUGAUUUCCAUCUA 717 15460 CDS 848 83.6%CCCCCCGGAGGUGAUUUCCAUCUAC 718 15461 CDS 851 72.2%CCCGGAGGUGAUUUCCAUCUACAAC 719 15462 CDS 853 85.8%CGGAGGUGAUUUCCAUCUACAACAG 720 15463 CDS 855 67.1%GAGGUGAUUUCCAUCUACAACAGCA 721 15464 CDS 952 68.9%ACUACGCCAAGGAGGUUUACAAAAU 722 15465 CDS 963 81.1%GAGGUUUACAAAAUAGACAUGCCGC 723 15466 CDS 1107 82.1%UUCUACAGACCCUACUUCAGAAUUG 724 15467 CDS 1108 99.1%UCUACAGACCCUACUUCAGAAUUGU 725 15468 CDS 1109 95.1%CUACAGACCCUACUUCAGAAUUGUU 726 15469 CDS 1129 90.4%UUGUUCGAUUUGACGUCUCAGCAAU 727 15470 CDS 1130 76.7%UGUUCGAUUUGACGUCUCAGCAAUG 728 15471 CDS 1131 79.7%GUUCGAUUUGACGUCUCAGCAAUGG 729 15472 CDS 1132 87.5%UUCGAUUUGACGUCUCAGCAAUGGA 730 15473 CDS 1144 66.9%UCUCAGCAAUGGAGAAGAAUGCUUC 731 15474 CDS 1145 76.6%CUCAGCAAUGGAGAAGAAUGCUUCC 732 15475 CDS 1147 88.9%CAGCAAUGGAGAAGAAUGCUUCCAA 733 15476 CDS 1162 84.5%AUGCUUCCAAUUUGGUGAAAGCAGA 734 15477 CDS 1163 89.2%UGCUUCCAAUUUGGUGAAAGCAGAG 735 15478 CDS 1165 86.6%CUUCCAAUUUGGUGAAAGCAGAGUU 736 15479 CDS 1177 61.2%UGAAAGCAGAGUUCAGAGUCUUUCG 737 15480 CDS 1185 92.6%GAGUUCAGAGUCUUUCGUUUGCAGA 738 15481 CDS 1219 99.6%CCAGAGUGCCUGAACAACGGAUUGA 739 15482 CDS 1224 94.0%GUGCCUGAACAACGGAUUGAGCUAU 740 15483 CDS 1225 88.1%UGCCUGAACAACGGAUUGAGCUAUA 741 15484 CDS 1228 59.3%CUGAACAACGGAUUGAGCUAUAUCA 742 15485 CDS 1229 77.5%UGAACAACGGAUUGAGCUAUAUCAG 743 15486 CDS 1230 61.5%GAACAACGGAUUGAGCUAUAUCAGA 744 15487 CDS 1233 84.5%CAACGGAUUGAGCUAUAUCAGAUUC 745 15488 CDS 1238 87.7%GAUUGAGCUAUAUCAGAUUCUCAAG 746 15489 CDS 1239 78.7%AUUGAGCUAUAUCAGAUUCUCAAGU 747 15490 CDS 1240 94.1%UUGAGCUAUAUCAGAUUCUCAAGUC 748 15491 CDS 1247 92.6%AUAUCAGAUUCUCAAGUCCAAAGAU 749 15492 CDS 1256 94.3%UCUCAAGUCCAAAGAUUUAACAUCU 750 15493 CDS 1259 99.1%CAAGUCCAAAGAUUUAACAUCUCCA 751 15494 CDS 1286 87.4%CCAGCGCUACAUCGACAGCAAAGUU 752 15495 CDS 1288 84.5%AGCGCUACAUCGACAGCAAAGUUGU 753 15496 CDS 1289 60.1%GCGCUACAUCGACAGCAAAGUUGUG 754 15497 CDS 1292 78.8%CUACAUCGACAGCAAAGUUGUGAAA 755 15498 CDS 1331 80.1%CGAAUGGCUCUCCUUCGAUGUAACU 756 15499 CDS 1353 62.4%ACUGAUGCUGUUCAUGAAUGGCUUC 757 15500 CDS 1361 74.3%UGUUCAUGAAUGGCUUCACCAUAAA 758 15501 CDS 1362 75.1%GUUCAUGAAUGGCUUCACCAUAAAG 759 15502 CDS 1363 87.2%UUCAUGAAUGGCUUCACCAUAAAGA 760 15503 CDS 1364 70.4%UCAUGAAUGGCUUCACCAUAAAGAC 761 15504 CDS 1365 100.7%CAUGAAUGGCUUCACCAUAAAGACA 762 15505 CDS 1368 100.1%GAAUGGCUUCACCAUAAAGACAGGA 763 15506 CDS 1398 92.0%GGAUUUAAAAUAAGCUUACACUGUC 764 15507 CDS 1399 83.2%GAUUUAAAAUAAGCUUACACUGUCC 765 15508 CDS 1415 85.6%ACACUGUCCCUGCUGCACUUUUGUA 766 15509 CDS 1418 97.4%CUGUCCCUGCUGCACUUUUGUACCA 767 15510 CDS 1420 59.1%GUCCCUGCUGCACUUUUGUACCAUC 768 15511 CDS 1421 73.7%UCCCUGCUGCACUUUUGUACCAUCU 769 15512 CDS 1422 79.5%CCCUGCUGCACUUUUGUACCAUCUA 770 15513 CDS 1451 62.7%UUACAUCAUCCCAAAUAAAAGUGAA 771 15514 CDS 1452 76.0%UACAUCAUCCCAAAUAAAAGUGAAG 772 15515 CDS 1470 44.7%AGUGAAGAACUAGAAGCAAGAUUUG 773 15516 CDS 1472 75.6%UGAAGAACUAGAAGCAAGAUUUGCA 774 15517 CDS 1474 96.8%AAGAACUAGAAGCAAGAUUUGCAGG 775 15518 CDS 1475 94.3%AGAACUAGAAGCAAGAUUUGCAGGU 776 15519 CDS 1476 63.3%GAACUAGAAGCAAGAUUUGCAGGUA 777 15520 CDS 1480 65.9%UAGAAGCAAGAUUUGCAGGUAUUGA 778 15521 CDS 1481 59.6%AGAAGCAAGAUUUGCAGGUAUUGAU 779 15522 CDS 1482 56.0%GAAGCAAGAUUUGCAGGUAUUGAUG 780 15523 CDS 1483 69.2%AAGCAAGAUUUGCAGGUAUUGAUGG 781 15524 CDS 1484 64.5%AGCAAGAUUUGCAGGUAUUGAUGGC 782 15525 CDS 1485 92.0%GCAAGAUUUGCAGGUAUUGAUGGCA 783 15526 CDS 1486 101.7%CAAGAUUUGCAGGUAUUGAUGGCAC 784 15527 CDS 1496 103.3%AGGUAUUGAUGGCACCUCCACAUAU 785 15528 CDS 1503 102.3%GAUGGCACCUCCACAUAUACCAGUG 786 15529 CDS 1506 86.6%GGCACCUCCACAUAUACCAGUGGUG 787 15530 CDS 1510 79.9%CCUCCACAUAUACCAGUGGUGAUCA 788 15531 CDS 1511 44.9%CUCCACAUAUACCAGUGGUGAUCAG 789 15532 CDS 1512 57.3%UCCACAUAUACCAGUGGUGAUCAGA 790 15533 CDS 1517 64.9%AUAUACCAGUGGUGAUCAGAAAACU 791 15534 CDS 1518 90.8%UAUACCAGUGGUGAUCAGAAAACUA 792 15535 CDS 1520 47.1%UACCAGUGGUGAUCAGAAAACUAUA 793 15536 CDS 1526 55.7%UGGUGAUCAGAAAACUAUAAAGUCC 794 15537 CDS 1527 89.6%GGUGAUCAGAAAACUAUAAAGUCCA 795 15538 CDS 1529 92.4%UGAUCAGAAAACUAUAAAGUCCACU 796 15539 CDS 1531 87.2%AUCAGAAAACUAUAAAGUCCACUAG 797 15540 CDS 1532 93.4%UCAGAAAACUAUAAAGUCCACUAGG 798 15541 CDS 1575 78.4%ACCCCACAUCUCCUGCUAAUGUUAU 799 15542 CDS 1576 84.6%CCCCACAUCUCCUGCUAAUGUUAUU 800 15543 CDS 1579 95.9%CACAUCUCCUGCUAAUGUUAUUGCC 801 15544 CDS 1591 89.6%UAAUGUUAUUGCCCUCCUACAGACU 802 15545 CDS 1592 85.0%AAUGUUAUUGCCCUCCUACAGACUU 803 15546 CDS 1598 51.2%AUUGCCCUCCUACAGACUUGAGUCA 804 15547 CDS 1650 39.4%GCUUUGGAUGCGGCCUAUUGCUUUA 805 15548 CDS 1652 82.3%UUUGGAUGCGGCCUAUUGCUUUAGA 806 15549 CDS 1653 86.1%UUGGAUGCGGCCUAUUGCUUUAGAA 807 15550 CDS 1655 80.0%GGAUGCGGCCUAUUGCUUUAGAAAU 808 15551 CDS 1657 72.3%AUGCGGCCUAUUGCUUUAGAAAUGU 809 15552 CDS 1658 72.2%UGCGGCCUAUUGCUUUAGAAAUGUG 810 15553 CDS 1659 57.8%GCGGCCUAUUGCUUUAGAAAUGUGC 811 15554 CDS 1660 83.4%CGGCCUAUUGCUUUAGAAAUGUGCA 812 15555 CDS 1662 79.3%GCCUAUUGCUUUAGAAAUGUGCAGG 813 15556 CDS 1663 86.3%CCUAUUGCUUUAGAAAUGUGCAGGA 814 15557 CDS 1664 84.8%CUAUUGCUUUAGAAAUGUGCAGGAU 815 15558 CDS 1665 71.1%UAUUGCUUUAGAAAUGUGCAGGAUA 816 15559 CDS 1666 61.8%AUUGCUUUAGAAAUGUGCAGGAUAA 817 15560 CDS 1667 84.9%UUGCUUUAGAAAUGUGCAGGAUAAU 818 15561 CDS 1668 82.8%UGCUUUAGAAAUGUGCAGGAUAAUU 819 15562 CDS 1670 69.8%CUUUAGAAAUGUGCAGGAUAAUUGC 820 15563 CDS 1671 90.2%UUUAGAAAUGUGCAGGAUAAUUGCU 821 15564 CDS 1672 68.6%UUAGAAAUGUGCAGGAUAAUUGCUG 822 15565 CDS 1678 74.2%AUGUGCAGGAUAAUUGCUGCCUACG 823 15566 CDS 1761 58.6%GGGUACAAUGCCAACUUCUGUGCUG 824 15567 CDS 1767 86.3%AAUGCCAACUUCUGUGCUGGAGCAU 825 15568 CDS 1782 83.7%GCUGGAGCAUGCCCGUAUUUAUGGA 826 15569 CDS 1783 86.9%CUGGAGCAUGCCCGUAUUUAUGGAG 827 15570 CDS 1786 90.5%GAGCAUGCCCGUAUUUAUGGAGUUC 828 15571 CDS 1787 91.1%AGCAUGCCCGUAUUUAUGGAGUUCA 829 15572 CDS 1788 68.0%GCAUGCCCGUAUUUAUGGAGUUCAG 830 15573 CDS 1789 75.7%CAUGCCCGUAUUUAUGGAGUUCAGA 831 15574 CDS 1796 88.9%GUAUUUAUGGAGUUCAGACACUCAG 832 15575 CDS 1800 52.5%UUAUGGAGUUCAGACACUCAGCACA 833 15576 CDS 1907 90.8%AACCAUUCUCUACUACAUUGGCAAA 834 15577 CDS 1924 70.2%UUGGCAAAACACCCAAGAUUGAACA 835 15578 CDS 1925 77.5%UGGCAAAACACCCAAGAUUGAACAG 836 15579 CDS/3UTR 1973 91.1%UUGCAAAUGCAGCUAAAAUUCUUGG 837 15580 3UTR 2020 70.1%CAAUGAUGAUGAUAAUGAUGAUGAC 838 15581 3UTR 2022 43.3%AUGAUGAUGAUAAUGAUGAUGACGA 839 15582 3UTR 2023 60.3%UGAUGAUGAUAAUGAUGAUGACGAC 840 15583 3UTR 2025 75.4%AUGAUGAUAAUGAUGAUGACGACGA 841 15584 3UTR 2026 40.8%UGAUGAUAAUGAUGAUGACGACGAC 842 15585 3UTR 2028 51.8%AUGAUAAUGAUGAUGACGACGACAA 843 15586 3UTR 2029 59.1%UGAUAAUGAUGAUGACGACGACAAC 844 15587 3UTR 2031 51.3%AUAAUGAUGAUGACGACGACAACGA 845 15588 3UTR 2032 32.7%UAAUGAUGAUGACGACGACAACGAU 846 15589 3UTR 2034 33.8%AUGAUGAUGACGACGACAACGAUGA 847 15590 3UTR 2035 57.0%UGAUGAUGACGACGACAACGAUGAU 848 15591 3UTR 2039 40.5%GAUGACGACGACAACGAUGAUGCUU 849 15592 3UTR 2045 56.8%GACGACAACGAUGAUGCUUGUAACA 850 15593 3UTR 2046 28.5%ACGACAACGAUGAUGCUUGUAACAA 851 15594 3UTR 2065 44.7%UAACAAGAAAACAUAAGAGAGCCUU 852 15595 3UTR 2066 58.3%AACAAGAAAACAUAAGAGAGCCUUG 853 15596 3UTR 2067 62.9%ACAAGAAAACAUAAGAGAGCCUUGG 854 15597 3UTR 2072 38.1%AAAACAUAAGAGAGCCUUGGUUCAU 855 15598 3UTR 2073 44.6%AAACAUAAGAGAGCCUUGGUUCAUC 856 15599 3UTR 2079 53.6%AAGAGAGCCUUGGUUCAUCAGUGUU 857 15600 3UTR 2081 33.2%GAGAGCCUUGGUUCAUCAGUGUUAA 858 15601 3UTR 2083 28.2%GAGCCUUGGUUCAUCAGUGUUAAAA 859 15602 3UTR 2110 46.5%UUUUUGAAAAGGCGGUACUAGUUCA 860 15603 3UTR 2116 56.1%AAAAGGCGGUACUAGUUCAGACACU 861 15604 3UTR 2117 60.9%AAAGGCGGUACUAGUUCAGACACUU 862 15605 3UTR 2136 76.8%ACACUUUGGAAGUUUGUGUUCUGUU 863 15606 3UTR 2137 29.5%CACUUUGGAAGUUUGUGUUCUGUUU 864 15607 3UTR 2140 62.6%UUUGGAAGUUUGUGUUCUGUUUGUU 865 15608 3UTR 2145 50.7%AAGUUUGUGUUCUGUUUGUUAAAAC 866 15609 3UTR 2147 62.9%GUUUGUGUUCUGUUUGUUAAAACUG 867 15610 3UTR 2148 59.7%UUUGUGUUCUGUUUGUUAAAACUGG 868 15611 3UTR 2149 50.3%UUGUGUUCUGUUUGUUAAAACUGGC 869 15612 3UTR 2150 49.8%UGUGUUCUGUUUGUUAAAACUGGCA 870 15613 3UTR 2152 55.2%UGUUCUGUUUGUUAAAACUGGCAUC 871 15614 3UTR 2153 82.2%GUUCUGUUUGUUAAAACUGGCAUCU 872 15615 3UTR 2154 70.0%UUCUGUUUGUUAAAACUGGCAUCUG 873 15616 3UTR 2155 45.5%UCUGUUUGUUAAAACUGGCAUCUGA 874 15617 3UTR 2156 54.9%CUGUUUGUUAAAACUGGCAUCUGAC 875 15618 3UTR 2189 40.4%AGUUGAAGGCCUUAUUCUACAUUUC 876 15619 3UTR 2190 34.1%GUUGAAGGCCUUAUUCUACAUUUCA 877 15620 3UTR 2207 91.3%ACAUUUCACCUACUUUGUAAGUGAG 878 15621 3UTR 2265 60.9%AAUAAACACUGGAAGAAUUUAUUAG 879 15622 3UTR 2267 36.4%UAAACACUGGAAGAAUUUAUUAGUG 880 15623 3UTR 2295 40.6%AUUAUGUGAACAACGACAACAACAA 881 15624 3UTR 2296 33.6%UUAUGUGAACAACGACAACAACAAC 882 15625 3UTR 2297 32.7%UAUGUGAACAACGACAACAACAACA 883 15626 3UTR 2298 40.8%AUGUGAACAACGACAACAACAACAA 884 15627 3UTR 2299 38.5%UGUGAACAACGACAACAACAACAAC 885 15628 3UTR 2301 84.2%UGAACAACGACAACAACAACAACAA 886 15629 3UTR 2302 43.2%GAACAACGACAACAACAACAACAAC 887 15630 3UTR 2304 57.8%ACAACGACAACAACAACAACAACAA 888 15631 3UTR 2305 44.3%CAACGACAACAACAACAACAACAAC 889 15632 3UTR 2308 38.7%CGACAACAACAACAACAACAACAAA 890 15633 3UTR 2309 37.4%GACAACAACAACAACAACAACAAAC 891 15634 3UTR 2314 73.5%CAACAACAACAACAACAAACAGGAA 892 15635 3UTR 2315 54.2%AACAACAACAACAACAAACAGGAAA 893 15636 3UTR 2389 30.7%CUUGAUUUUUCUGUAUUGCUAUGCA 894 15637 3UTR 2435 16.0%ACUCUUAGAGUUAACAGUGAGUUAU 895 15638 3UTR 2445 18.4%UUAACAGUGAGUUAUUUAUUGUGUG 896 15639 3UTR 2471 36.3%UACUAUAUAAUGAACGUUUCAUUGC 897 15640 3UTR 2472 73.3%ACUAUAUAAUGAACGUUUCAUUGCC 898 15641 3UTR 2484 63.4%ACGUUUCAUUGCCCUUGGAAAAUAA 899 15642 3UTR 2488 65.4%UUCAUUGCCCUUGGAAAAUAAAACA 900 15643 3UTR 2493 39.3%UGCCCUUGGAAAAUAAAACAGGUGU 901 15644 3UTR 2519 66.7%UAAAGUGGAGACCAAAUACUUUGCC 902 15645 3UTR 2520 40.1%AAAGUGGAGACCAAAUACUUUGCCA 903 15646 3UTR 2526 40.9%GAGACCAAAUACUUUGCCAGAAACU 904 15647 3UTR 2527 41.5%AGACCAAAUACUUUGCCAGAAACUC 905 15648 3UTR 2528 47.6%GACCAAAUACUUUGCCAGAAACUCA 906 15649 3UTR 2529 47.6%ACCAAAUACUUUGCCAGAAACUCAU 907 15650 3UTR 2530 31.9%CCAAAUACUUUGCCAGAAACUCAUG 908 15651 3UTR 2531 29.0%CAAAUACUUUGCCAGAAACUCAUGG 909 15652 3UTR 2537 78.0%CUUUGCCAGAAACUCAUGGAUGGCU 910 15653 3UTR 2538 52.4%UUUGCCAGAAACUCAUGGAUGGCUU 911 15654 3UTR 2540 59.7%UGCCAGAAACUCAUGGAUGGCUUAA 912 15655 3UTR 2541 45.1%GCCAGAAACUCAUGGAUGGCUUAAG 913 15656 3UTR 2542 42.1%CCAGAAACUCAUGGAUGGCUUAAGG 914 15657 3UTR 2543 76.9%CAGAAACUCAUGGAUGGCUUAAGGA 915 15658 3UTR 2544 29.0%AGAAACUCAUGGAUGGCUUAAGGAA 916 15659 3UTR 2547 45.2%AACUCAUGGAUGGCUUAAGGAACUU 917 15660 3UTR 2560 38.4%CUUAAGGAACUUGAACUCAAACGAG 918 15661 3UTR 2561 33.3%UUAAGGAACUUGAACUCAAACGAGC 919 15662 3UTR 2562 31.9%UAAGGAACUUGAACUCAAACGAGCC 920 15663 3UTR 2563 44.5%AAGGAACUUGAACUCAAACGAGCCA 921 15664 3UTR 2564 90.1%AGGAACUUGAACUCAAACGAGCCAG 922 15665 3UTR 2566 64.4%GAACUUGAACUCAAACGAGCCAGAA 923 15666 3UTR 2623 32.5%AAGUGAGUUAUUAUAUGACCGAGAA 924 15667 3UTR 2681 34.0%UGUUAUGUAUCAGCUGCCUAAGGAA 925 15668 3UTR 2791 59.0%UUUAAUUGUAAAUGGUUCUUUGUCA 926 15669 3UTR 2792 56.3%UUAAUUGUAAAUGGUUCUUUGUCAG 927 15670 3UTR 2793 46.8%UAAUUGUAAAUGGUUCUUUGUCAGU 928 15671 3UTR 2795 53.2%AUUGUAAAUGGUUCUUUGUCAGUUU 929 15672 3UTR 2798 33.1%GUAAAUGGUUCUUUGUCAGUUUAGU 930 15673 3UTR 2809 32.8%UUUGUCAGUUUAGUAAACCAGUGAA 931 15674 3UTR 2813 40.9%UCAGUUUAGUAAACCAGUGAAAUGU 932 15675 3UTR 2816 38.1%GUUUAGUAAACCAGUGAAAUGUUGA 933 15676 3UTR 2840 59.4%AAAUGUUUUGACAUGUACUGGUCAA 934 15677 3UTR 2957 77.9%UGGAUAUAGAAGCCAGCAUAAUUGA 935 15678 3UTR 2958 74.1%GGAUAUAGAAGCCAGCAUAAUUGAA 936 15679 3UTR 2959 52.4%GAUAUAGAAGCCAGCAUAAUUGAAA 937 15680 3UTR 2963 49.9%UAGAAGCCAGCAUAAUUGAAAACAC 938 15681 3UTR 2964 45.3%AGAAGCCAGCAUAAUUGAAAACACA 939 15682 3UTR 2966 45.5%AAGCCAGCAUAAUUGAAAACACAUC 940 15683 3UTR 3137 60.5%ACAAAUGUAUGUUUCUUUUAGCUGG 941 15684 3UTR 3138 63.6%CAAAUGUAUGUUUCUUUUAGCUGGC 942 15685 3UTR 3142 58.4%UGUAUGUUUCUUUUAGCUGGCCAGU 943 15686 3UTR 3144 56.3%UAUGUUUCUUUUAGCUGGCCAGUAC 944 15687 3UTR 3145 52.1%AUGUUUCUUUUAGCUGGCCAGUACU 945 15688 3UTR 3147 74.6%GUUUCUUUUAGCUGGCCAGUACUUU 946 15689 3UTR 3150 70.4%UCUUUUAGCUGGCCAGUACUUUUGA 947 15690 3UTR 3154 61.7%UUAGCUGGCCAGUACUUUUGAGUAA 948 15691 3UTR 3156 52.3%AGCUGGCCAGUACUUUUGAGUAAAG 949 15692 3UTR 3157 72.2%GCUGGCCAGUACUUUUGAGUAAAGC 950 15693 3UTR 3158 62.4%CUGGCCAGUACUUUUGAGUAAAGCC 951 15694 3UTR 3180 49.0%GCCCCUAUAGUUUGACUUGCACUAC 952 15695 3UTR 3182 43.9%CCCUAUAGUUUGACUUGCACUACAA 953 15696 3UTR 3183 35.2%CCUAUAGUUUGACUUGCACUACAAA 954 15697 3UTR 3184 38.1%CUAUAGUUUGACUUGCACUACAAAU 955 15698 3UTR 3185 73.3%UAUAGUUUGACUUGCACUACAAAUG 956 15699 3UTR 3256 86.3%UUCAUUAUUAUGACAUAAGCUACCU 957 15700 3UTR 3258 61.6%CAUUAUUAUGACAUAAGCUACCUGG 958 15701 3UTR 3342 66.0%UUCAUCUUCCAAGCAUCAUUACUAA 959 15702 3UTR 3346 67.3%UCUUCCAAGCAUCAUUACUAACCAA 960 15703 3UTR 3358 63.6%CAUUACUAACCAAGUCAGACGUUAA 961 15704 3UTR 3396 71.8%UAGGAAAAGGAGGAAUGUUAUAGAU 962 15705 3UTR 3550 69.1%UUGUUAUUACAAAGAGGACACUUCA 963 15706 3UTR 3657 72.3%GGGGAAAAAAGUCCAGGUCAGCAUA 964 15707 3UTR 3671 79.7%AGGUCAGCAUAAGUCAUUUUGUGUA 965 15708 3UTR 3779 57.5%UUUCUUUCCUCUGAGUGAGAGUUAU 966 15709 3UTR 3783 62.6%UUUCCUCUGAGUGAGAGUUAUCUAU 967 15710 3UTR 3932 61.3%UAAAAAUUAAUAGGCAAAGCAAUGG 968 15711 3UTR 3934 44.3%AAAAUUAAUAGGCAAAGCAAUGGAA 969 15712 3UTR 4034 68.7%UUUUUUGGAAUUUCCUGACCAUUAA 970 15713 3UTR 4058 50.6%AUUAAAGAAUUGGAUUUGCAAGUUU 971 15714 3UTR 4120 69.8%UAAACAGCCCUUGUGUUGGAUGUAA 972 15715 3UTR 4147 39.5%CAAUCCCAGAUUUGAGUGUGUGUUG 973 15716 3UTR 4148 62.2%AAUCCCAGAUUUGAGUGUGUGUUGA 974 15717 3UTR 4152 34.2%CCAGAUUUGAGUGUGUGUUGAUUAU 975 15718 3UTR 4273 38.0%GUCUUUCCUCAUGAAUGCACUGAUA 976 15719 3UTR 4460 48.5%UAUUUUUGUGUUAAUCAGCAGUACA 977 15720 3UTR 4482 37.1%ACAAUUUGAUCGUUGGCAUGGUUAA 978 15721 3UTR 4580 60.1%GUUUUUGUGGUGCUCUAGUGGUAAA 979 15722 3UTR 4583 50.6%UUUGUGGUGCUCUAGUGGUAAAUAA 980 15723 3UTR 4584 42.1%UUGUGGUGCUCUAGUGGUAAAUAAA 981 15724 3UTR 4642 91.3%UCAGUACCAUCAUCGAGUCUAGAAA 982 15725 3UTR 4737 90.4%UUCUCCCUUAAGGACAGUCACUUCA 983 15726 3UTR 4751 94.6%CAGUCACUUCAGAAGUCAUGCUUUA 984 15727 3UTR 4753 87.2%GUCACUUCAGAAGUCAUGCUUUAAA 985 15728 3UTR 4858 70.2%GUAAUUGUUUGAGAUUUAGUUUCCA 986 15729 3UTR 4963 81.2%CGCCAGGGCCAAAAGAACUGGUCUA 987 15730 3UTR 5177 81.4%CCAGACUCCUCAAACGAGUUGCCAA 988

TABLE 7 hTGFB2 sd-rxRNA Target Gene hTGFB2 SEQ ID Duplex ID SingleStrand ID sd-rxRNA sequences NO 18570 17560mU.A.mU.mU.mU.A.mU.mU.G.mU.G.mU.A.Chl 989 17562P.mU.A.fC.A.fC.A.A.fU.A.A.A.fU.A*A*fC*fU*fC*A*C 990 18571 17561mU.mU.A.mU.mU.mU.A.mU.mU.G.mU.G.mU.A.Chl 991 17562P.mU.A.fC.A.fC.A.A.fU.A.A.A.fU.A*A*fC*fU*fC*A*C 992 18572 17563A.mU.mC.A.G.mU.G.mU.mU.A.A.A.A.Chl 993 17565P.mU.fU.fU.fU.A.A.fC.A.fC.fU.G.A.fU*G*A*A*fC*fC*A 994 18573 17564mC.A.mU.mC.A.G.mU.G.mU.mU.A.A.A.A.Chl 995 17565P.mU.fU.fU.fU.A.A.fC.A.fC.fU.G.A.fU*G*A*A*fC*fC*A 996 18574 17566A.mU.G.G.mC.mU.mU.A.A.G.G.A.A.Chl 997 17568P.mU.fU.fC.fC.fU.fU.A.A.G.fC.fC.A.fU*fC*fC*A*fU*G*A 998 18575 17567G.A.mU.G.G.mC.mU.mU.A.A.G.G.A.A.Chl 999 17568P.mU.fU.fC.fC.fU.fU.A.A.G.fC.fC.A.fU*fC*fC*A*fU*G*A 1000 18576 17569mU.mU.G.mU.G.mU.mU.mC.mU.G.mU.mU.A.Chl 1001 17571P.mU.A.A.fC.A.G.A.A.fC.A.fC.A.A*A*fC*fU*fU*fC*C 1002 18577 17570mU.mU.mU.G.mU.G.mU.mU.mC.mU.G.mU.mU.A.Chl 1003 17571P.mU.A.A.fC.A.G.A.A.fC.A.fC.A.A*A*fC*fU*fU*fC*C 1004 18578 17572A.A.A.mU.A.mC.mU.mU.mU.G.mC.mC.A.Chl 1005 17574P.mU.G.G.fC.A.A.A.G.fU.A.fU.fU.fU*G*G*fU*fC*fU*C 1006 18579 17573mC.A.A.A.mU.A.mC.mU.mU.mU.G.mC.mC.A.Chl 1007 17574P.mU.G.G.fC.A.A.A.G.fU.A.fU.fU.fU*G*G*fU*fC*fU*C 1008 18580 17575mC.mU.mU.G.mC.A.mC.mU.A.mC.A.A.A.Chl 1009 17577P.mU.fU.fU.G.fU.A.G.fU.G.fC.A.A.G*fU*fC*A*A*A*C 1010 18581 17576A.mC.mU.mU.G.mC.A.mC.mU.A.mC.A.A.A.Chl 1011 17577P.mU.fU.fU.G.fU.A.G.fU.G.fC.A.A.G*fU*fC*A*A*A*C 1012 18582 17578G.A.A.mU.mU.mU.A.mU.mU.A.G.mU.A.Chl 1013 17580P.mU.A.fC.fU.A.A.fU.A.A.A.fU.fU.fC*fU*fU*fC*fC*A*G 1014 18583 17579A.G.A.A.mU.mU.mU.A.mU.mU.A.G.mU.A.Chl 1015 17580P.mU.A.fC.fU.A.A.fU.A.A.A.fU.fU.fC*fU*fU*fC*fC*A*G 1016 18584 17581mU.mU.G.mC.A.mC.mU.A.mC.A.A.A.A.Chl 1017 17583P.mU.fU.fU.fU.G.fU.A.G.fU.G.fC.A.A*G*fU*fC*A*A*A 1018 18585 17582mC.mU.mU.G.mC.A.mC.mU.A.mC.A.A.A.A.Chl 1019 17583P.mU.fU.fU.fU.G.fU.A.G.fU.G.fC.A.A*G*fU*fC*A*A*A 1020 18586 17584A.mU.A.A.A.A.mC.A.G.G.mU.G.A.Chl 1021 17586P.mU.fC.A.fC.fC.fU.G.fU.fU.fU.fU.A.fU*fU*fU*fU*fC*fC*A 1022 18587 17585A.A.mU.A.A.A.A.mC.A.G.G.mU.G.A.Chl 1023 17586P.mU.fC.A.fC.fC.fU.G.fU.fU.fU.fU.A.fU*fU*fU*fU*fC*fC*A 1024 18588 17587G.A.mC.A.A.mC.A.A.mC.A.A.mC.A.Chl 1025 17588P.mU.G.fU.fU.G.fU.fU.G.fU.fU.G.fU.fC*G*fU*fU*G*fU*U 1026 18589 17589A.mU.G.mC.mU.mU.G.mU.A.A.mC.A.A.Chl 1027 17590P.mU.fU.G.fU.fU.A.fC.A.A.G.fC.A.fU*fC*A*fU*fC*G*U 1028 18590 17591mC.A.G.A.A.A.mC.mU.mC.A.mU.G.A.Chl 1029 17592P.mU.fC.A.fU.G.A.G.fU.fU.fU.fC.fU.G*G*fC*A*A*A*G 1030 18591 17593G.mU.A.mU.mU.G.mC.mU.A.mU.G.mC.A.Chl 1031 17594P.mU.G.fC.A.fU.A.G.fC.A.A.fU.A.fC*A*G*A*A*A*A 1032 18592 17595mC.mC.A.G.A.A.A.mC.mU.mC.A.mU.A.Chl 1033 17596P.mU.A.fU.G.A.G.fU.fU.fU.fC.fU.G.G*fC*A*A*A*G*U 1034 18593 17597A.mC.mU.mC.A.A.A.mC.G.A.G.mC.A.Chl 1035 17598P.mU.G.fC.fU.fC.G.fU.fU.fU.G.A.G.fU*fU*fC*A*A*G*U 1036 18594 17599A.mU.A.mU.G.A.mC.mC.G.A.G.A.A.Chl 1037 17600P.mU.fU.fC.fU.fC.G.G.fU.fC.A.fU.A.fU*A*A*fU*A*A*C 1038 18595 17601mC.G.A.mC.G.A.mC.A.A.mC.G.A.A.Chl 1039 17602P.mU.fU.fC.G.fU.fU.G.fU.fC.G.fU.fC.G*fU*fC*A*fU*fC*A 1040 18596 17603G.mU.A.A.A.mC.mC.A.G.mU.G.A.A.Chl 1041 17604P.mU.fU.fC.A.fC.fU.G.G.fU.fU.fU.A.fC*fU*A*A*A*fC*U 1042 18597 17605mU.mU.G.mU.mC.A.G.mU.mU.mU.A.G.A.Chl 1043 17606P.mU.fC.fU.A.A.A.fC.fU.G.A.fC.A.A*A*G*A*A*fC*C 1044 18598 17607mU.mC.A.mU.mC.A.G.mU.G.mU.mU.A.A.Chl 1045 17608P.mU.fU.A.A.fC.A.fC.fU.G.A.fU.G.A*A*fC*fC*A*A*G 1046 18599 17609A.A.mC.mU.mC.A.A.A.mC.G.A.G.A.Chl 1047 17610P.mU.fC.fU.fC.G.fU.fU.fU.G.A.G.fU.fU*fC*A*A*G*fU*U 1048 18600 17611mC.G.A.mC.A.A.mC.A.A.mC.A.A.A.Chl 1049 17612P.mU.fU.fU.G.fU.fU.G.fU.fU.G.fU.fC.G*fU*fU*G*fU*fU*C 1050 18601 17613A.mC.G.A.mC.A.A.mC.G.A.mU.G.A.Chl 1051 17614P.mU.fC.A.fU.fC.G.fU.fU.G.fU.fC.G.fU*fC*G*fU*fC*A*U 1052 18602 17615G.mC.mU.G.mC.mC.mU.A.A.G.G.A.A.Chl 1053 17616P.mU.fU.fC.fC.fU.fU.A.G.G.fC.A.G.fC*fU*G*A*fU*A*C 1054 18603 17617A.mU.mU.mC.mU.A.mC.A.mU.mU.mU.mC.A.Chl 1055 17618P.mU.G.A.A.A.fU.G.fU.A.G.A.A.fU*A*A*G*G*fC*C 1056 18604 17619G.mU.G.mU.G.mU.mU.G.A.mU.mU.A.A.Chl 1057 17620P.mU.fU.A.A.fU.fC.A.A.fC.A.fC.A.fC*A*fC*fU*fC*A*A 1058 Rat TGFB2sd-rxRNA 18678 18604 mC.G.G.mU.G.A.mC.A.A.mU.G.A.A-chol 1061 18605mU.fU.fC.A.fU.fU.G.fU.fC.A.fC.fC.G*fU*G*A*fU*fU*U 1062 18679 18606mU.mU.G.mU.mC.mU.mC.G.G.mU.A.mU.A-chol 1063 18607mU.A.fU.A.fC.fC.G.A.G.A.fC.A.A*A*G*G*G*A*A 1064 18680 18608G.A.G.mU.mU.G.mU.A.mU.G.mU.A.A-chol 1065 18609mU.fU.A.fC.A.fU.A.fC.A.A.fC.fU.fC*fC*A*fC*fU*G*A 1066 18681 18610A.mU.mU.mU.G.mU.mU.A.G.mU.G.mU.A-chol 1067 18611mU.A.fC.A.fC.fU.A.A.fC.A.A.A.fU*fU*fC*fU*fU*fC*C 1068 18682 18612G.mC.A.A.G.mU.mC.mU.G.A.G.A.A-chol 1069 18613mU.fU.fC.fU.fC.A.G.A.fC.fU.fU.G.fC*fU*fC*A*G*fU*U 1070 18683 18614A.A.A.mU.mC.A.mC.G.G.mU.G.A.A-chol 1071 18615mU.fU.fC.A.fC.fC.G.fU.G.A.fU.fU.fU*fU*fC*A*fU*fC*C 1072 18684 18616A.A.A.mU.G.mC.A.G.mC.mU.A.A.A-chol 1073 18617mU.fU.fU.A.G.fC.fU.G.fC.A.fU.fU.fU*A*fC*A*A*G*A 1074 18685 18618mC.mU.mU.G.G.A.A.A.A.mU.A.A.A-chol 1075 18619mU.fU.fU.A.fU.fU.fU.fU.fC.fC.A.A.G*G*G*fC*A*A*U 1076 18686 18620mC.mC.mU.mU.mU.G.A.A.mU.A.A.A.A-chol 1077 18621mU.fU.fU.fU.A.fU.fU.fC.A.A.A.G.G*fU*A*fC*fU*G*G 1078 18687 18622A.A.mC.A.mC.A.mC.mU.G.mC.A.A.A-chol 1079 18623mU.fU.fU.G.fC.A.G.fU.G.fU.G.fU.fU*fU*fU*fC*A*fU*C 1080 18688 18624A.A.A.A.mC.A.mC.A.mC.mU.G.mC.A-chol 1081 18625mU.G.fC.A.G.fU.G.fU.G.fU.fU.fU.fU*fC*A*fU*fC*A*U 1082 18689 18626G.A.A.G.G.mC.mC.mU.G.mU.mU.A.A-chol 1083 18627mU.fU.A.A.fC.A.G.G.fC.fC.fU.fU.fC*fU*G*G*A*fC*A 1084 18690 18628mU.A.mU.mU.G.mC.mU.mC.mU.G.mC.A.A-chol 1085 18629mU.fU.G.fC.A.G.A.G.fC.A.A.fU.A*fC*A*G*A*G*G 1086

TABLE 8 hSPP1 sd-rxRNA Target Gene hSPP1 SEQ ID Duplex ID Single StrandID sd-rxRNA sequence NO 18538 17430 G.A.mU.G.A.A.mU.mC.mU.G.A.mU.A.Chl1087 17432 P.mU.A.fU.fC.A.G.A.fU.fU.fC.A.fU.fC*A*G*A*A*fU*G 1088 1853917431 mU.G.A.mU.G.A.A.mU.mC.mU.G.A.mU.A.Chl 1089 17432P.mU.A.fU.fC.A.G.A.fU.fU.fC.A.fU.fC*A*G*A*A*fU*G 1090 18540 17433A.mU.mU.mU.G.mC.mU.mU.mU.mU.G.mC.A.Chl 1091 17435P.mU.G.fC.A.A.A.A.G.fC.A.A.A.fU*fC*A*fC*fU*G*fC 1092 18541 17434G.A.mU.mU.mU.G.mC.mU.mU.mU.mU.G.mC.A.Chl 1093 17435P.mU.G.fC.A.A.A.A.G.fC.A.A.A.fU*fC*A*fC*fU*G*fC 1094 18542 17436G.mU.G.A.mU.mU.mU.G.mC.mU.mU.mU.AChl 1095 17438P.mU.A.A.A.G.fC.A.A.A.fU.fC.A.fC*fU*G*fC*A*A*fU 1096 18543 17437A.G.mU.G.A.mU.mU.mU.G.mC.mU.mU.mU.A.Chl 1097 17438P.mU.A.A.A.G.fC.A.A.A.fU.fC.A.fC*fU*G*fC*A*A*fU 1098 18544 17439A.A.mU.mU.mU.mC.G.mU.A.mU.mU.mU.A.Chl 1099 17441P.mU.A.A.A.fU.A.fC.G.A.A.A.fU.fU*fU*fC*A*G*G*fU 1100 18545 17440A.A.A.mU.mU.mU.mC.G.mU.A.mU.mU.mU.A.Chl 1101 17441P.mU.A.A.A.fU.A.fC.G.A.A.A.fU.fU*fU*fC*A*G*G*fU 1102 18546 17442mC.A.mC.A.G.mC.mC.A.mU.G.A.A.A.Chl 1103 17444P.mU.fU.fU.C.A.fU.G.G.fC.fU.G.fU.G*A*A*A*fU*fU*fC 1104 18547 17443mU.mC.A.mC.A.G.mC.mC.A.mU.G.A.A.A.Chl 1105 17444P.mU.fU.fU.C.A.fU.G.G.fC.fU.G.fU.G*A*A*A*fU*fU*fC 1106 18548 17445G.A.mU.mU.mU.G.mC.mU.mU.mU.mU.G.A.Chl 1107 17447P.mU.fC.A.A.A.A.G.fC.A.A.A.fU.fC*A*fC*fU*G*fC*A 1108 18549 17446mU.G.A.mU.mU.mU.G.mC.mU.mU.mU.mU.G.A.Chl 1109 17447P.mU.fC.A.A.A.A.G.fC.A.A.A.fU.fC*A*fC*fU*G*fC*A 1110 18550 17448mU.mU.G.mC.mU.mU.mU.mU.G.mC.mC.mU.A.Chl 1111 17450P.mU.A.G.G.fC.A.A.A.A.G.fC.A.A*A*fU*fC*A*fC*U 1112 18551 17449mU.mU.mU.G.mC.mU.mU.mU.mU.G.mC.mC.mU.A.Chl 1113 17450P.mU.A.G.G.fC.A.A.A.A.G.fC.A.A*A*fU*fC*A*fC*U 1114 18552 17451mU.mU.mU.mC.mU.mC.A.G.mU.mU.mU.A.A.Chl 1115 17452P.mU.fU.A.A.A.fC.fU.G.A.G.A.A.A*G*A*A*G*fC*A 1116 18553 17453mU.mU.G.mC.A.mU.mU.mU.A.G.mU.mC.A.Chl 1117 17454P.mU.G.A.fC.fU.A.A.A.fU.G.fC.A.A*A*G*fU*G*A*G 1118 18554 17455A.mC.mU.mU.mU.G.mC.A.mU.mU.mU.A.A.Chl 1119 17456P.mU.fU.A.A.A.fU.G.fC.A.A.A.G.fU*G*A*G*A*A*A 1120 18555 17457A.mU.mU.mU.A.G.mU.mC.A.A.A.A.A.Chl 1121 17458P.mU.fU.fU.fU.fU.G.A.fC.fU.A.A.A.fU*G*fC*A*A*A*G 1122 18556 17459mU.mU.mC.mU.mU.mU.mC.mU.mC.A.G.mU.A.Chl 1123 17460P.mU.A.fC.fU.G.A.G.A.A.A.G.A.A*G*fC*A*fU*fU*fU 1124 18557 17461mU.mC.mU.mU.mU.mC.mU.mC.A.G.mU.mU.A.Chl 1125 17462P.mU.A.A.fC.fU.G.A.G.A.A.A.G.A*A*G*fC*A*fU*fU 1126 18558 17463G.A.A.A.G.A.G.A.A.mC.A.mU.A.Chl 1127 17464P.mU.A.fU.G.fU.fU.fC.fU.fC.fU.fU.fU.fC*A*fU*fU*fU*fU*G 1128 18559 17465mC.mU.mU.mU.G.mC.A.mU.mU.mU.A.G.A.Chl 1129 17466P.mU.fC.fU.A.A.A.fU.G.fC.A.A.A.G*fU*G*A*G*A*A 1130 18560 17467mU.mU.mU.G.mC.A.mU.mU.mU.A.G.mU.A.Chl 1131 17468P.mU.A.fC.fU.A.A.A.fU.G.fC.A.A.A*G*fU*G*A*G*A 1132 18561 17469mC.mU.mC.A.mC.mU.mU.mU.G.mC.A.mU.A.Chl 1133 17470P.mU.A.fU.G.fC.A.A.A.G.fU.G.A.G*A*A*A*fU*fU*G 1134 18562 17471mU.mU.mC.mU.mC.A.mC.mU.mU.mU.G.mC.A.Chl 1135 17472P.mU.G.fC.A.A.A.G.fU.G.A.G.A.A*A*fU*fU*G*fU*A 1136 18563 17473mC.A.mC.mU.mC.mC.A.G.mU.mU.G.mU.A.Chl 1137 17474P.mU.A.fC.A.A.fC.fU.G.G.A.G.fU.G*A*A*A*A*fC*U 1138 18564 17475A.A.mU.G.A.A.A.G.A.G.A.A.A.Chl 1139 17476P.mU.fU.fU.fC.fU.fC.fU.fU.fU.fC.A.fU.fU*fU*fU*G*fC*fU*A 1140 18565 17477mU.G.mC.A.G.mU.G.A.mU.mU.mU.mG.A.Chl 1141 17478P.mU.fC.A.A.A.fU.fC.A.fC.fU.G.fC.A*A*fU*fU*fC*fU*C 1142 18566 17479mU.G.A.A.A.G.A.G.A.A.mC.A.A.Chl 1143 17480P.mU.fU.G.fU.fU.fC.fU.fC.fU.fU.fU.fC.A*fU*fU*fU*fU*G*C 1144 18567 17481A.mC.mC.mU.G.A.A.A.mU.mU.mU.mC.A.Chl 1145 17482P.mU.G.A.A.A.fU.fU.fU.fC.A.G.G.fU*G*fU*fU*fU*A*U 1146 18568 17483G.A.A.mU.mU.G.mC.A.G.mU.G.A.A.Chl 1147 17484P.mU.fU.fC.A.fC.fU.G.fC.A.A.fU.fU.fC*fU*fC*A*fU*G*G 1148 18569 17485G.G.mC.mU.G.A.mU.mU.mC.mU.G.G.A.Chl 1149 17486P.mU.fC.fC.A.G.A.A.fU.fC.A.G.fC.fC*fU*G*fU*fU*fU*A 1150 Rat TargetingSPP1 18662 18630 G.mU.mU.mC.G.mU.mU.G.mU.mU.mU.mC.A-chol 1151 18631P.mU.G.A.A.A.fC.A.A.fC.G.A.A.fC*fU*A*A*G*fC*U 1152 18663 18632G.A.A.A.G.A.A.A.mU.A.G.A.A-chol 1153 18633P.mU.fU.fC.fU.A.fU.fU.fU.fC.fU.fU.fU.fC*fU*fC*fC*A*fC*A 1154 18664 18634G.mU.G.G.A.G.A.A.A.G.A.A.A-chol 1155 18635P.mU.fU.fU.fC.fU.fU.fU.fC.fU.fC.fC.A.fC*A*fU*A*fC*A*U 1156 18665 18636mC.mU.G.mU.G.mU.mC.A.mC.mU.A.mU.A-chol 1157 18637P.mU.A.fU.A.G.fU.G.A.fC.A.fC.A.G*A*fC*fU*A*fU*U 1158 18666 18638G.mU.mU.mU.mC.mU.mC.A.G.mU.mU.mC.A-chol 1159 18639P.mU.G.A.A.fC.fU.G.A.G.A.A.A.fC*A*A*G*fC*A*G 1160 18667 18640mU.A.mC.A.G.G.A.A.mC.A.G.mC.A-chol 1161 18641P.mU.G.fC.fU.G.fU.fU.fC.fC.fU.G.fU.A*A*G*fU*fU*fU*G 1162 18668 18642G.mC.A.G.G.mC.A.A.A.mC.mU.mU.A-chol 1163 18643P.mU.A.A.G.fU.fU.fU.G.fC.fC.fU.G.fC*fC*fU*fC*fU*A*C 1164 18669 18644A.A.mC.mU.mU.A.mC.A.G.G.A.A.A-chol 1165 18645P.mU.fU.fU.fC.fC.fU.G.fU.A.A.G.fU.fU*fU*G*fC*fC*fU*G 1166 18670 18646mC.A.mC.mU.G.mC.A.mU.mU.mU.mU.A.A-chol 1167 18647P.mU.fU.A.A.A.A.fU.G.fC.A.G.fU.G*G*fC*fC*A*fU*U 1168 18671 18648G.A.mC.A.mC.mC.A.mC.mU.G.mU.A.A-chol 1169 18649P.mU.fU.A.fC.A.G.fU.G.G.fU.G.fU.fC*fU*G*fC*A*fU*G 1170 18672 18650A.G.A.G.G.mC.A.G.G.mC.A.A.A-chol 1171 18651P.mU.fU.fU.G.fC.fC.fU.G.fC.fC.fU.fC.fU*A*fC*A*fU*A*C 1172 18673 18652mU.A.G.A.G.G.mC.A.G.G.mC.A.A-chol 1173 18653P.mU.fU.G.fC.fC.fU.G.fC.fC.fU.fC.fU.A*fC*A*fU*A*fC*A 1174 18674 18654G.A.G.A.G.mU.mU.mC.A.mU.mC.mU.A-chol 1175 18655P.mU.A.G.A.fU.G.A.A.fC.fU.fC.fU.fC*fU*A*A*fU*fU*C 1176 18675 18656mU.G.mU.G.A.A.mU.A.A.A.mU.mC.A-chol 1177 18657P.mU.G.A.fU.fU.fU.A.fU.U.fC.A.fC.A*fC*fC*A*fC*A*A 1178 18676 18658G.mU.G.A.A.mU.A.A.A.mU.mC.mU.A-chol 1179 18659P.mU.A.G.A.fU.fU.fU.A.fU.fU.fC.A.fC*A*fC*fC*A*fC*A 1180 18677 18660mU.G.A.AmU.A.A.A.mU.mC.mU.mU.A-chol 1181 18661P.mU.A.A.G.A.fU.fU.fU.A.fU.U.fC.A*fC*A*fC*fC*A*C 1182

TABLE 9 Inhibition of gene expression with hSPP1 ori sequences TargetGene Gene SEQ ID hSPP1 A549 0.1 nM Duplex ID Region Ref Pos NO SenseStrand Sequence Activity 14840 5UTR/CDS 155 1183AAGGAAAACUCACUACCAUGAGAAA  4.4% 14841 5UTR/CDS 161 1184AACUCACUACCAUGAGAAUUGCAGA 2.46% 14842 5UTR/CDS 163 1185CUCACUACCAUGAGAAUUGCAGUGA 20.54%  14843 5UTR/CDS 164 1186UCACUACCAUGAGAAUUGCAGUGAA  2.8% 14844 CDS 168 1187UACCAUGAGAAUUGCAGUGAUUUGA  3.6% 14845 CDS 169 1188ACCAUGAGAAUUGCAGUGAUUUGCA  5.2% 14846 CDS 171 1189CAUGAGAAUUGCAGUGAUUUGCUUA  0.8% 14847 CDS 172 1190AUGAGAAUUGCAGUGAUUUGCUUUA 0.95% 14848 CDS 173 1191UGAGAAUUGCAGUGAUUUGCUUUUA  3.2% 14849 CDS 174 1192GAGAAUUGCAGUGAUUUGCUUUUGA 4.14% 14850 CDS 175 1193AGAAUUGCAGUGAUUUGCUUUUGCA  2.9% 14851 CDS 176 1194GAAUUGCAGUGAUUUGCUUUUGCCA 8.38% 14852 CDS 177 1195AAUUGCAGUGAUUUGCUUUUGCCUA  4.6% 14853 CDS 180 1196UGCAGUGAUUUGCUUUUGCCUCCUA 11.1% 14854 CDS 181 1197GCAGUGAUUUGCUUUUGCCUCCUAA 10.87%  14855 CDS 182 1198CAGUGAUUUGCUUUUGCCUCCUAGA  5.3% 14856 CDS 206 1199GCAUCACCUGUGCCAUACCAGUUAA 15.29%  14857 CDS 208 1200AUCACCUGUGCCAUACCAGUUAAAA 22.6% 14858 CDS 212 1201CCUGUGCCAUACCAGUUAAACAGGA 13.3% 14859 CDS 215 1202GUGCCAUACCAGUUAAACAGGCUGA 21.2% 14860 CDS 216 1203UGCCAUACCAGUUAAACAGGCUGAA 20.24%  14861 CDS 220 1204AUACCAGUUAAACAGGCUGAUUCUA 12.5% 14862 CDS 221 1205UACCAGUUAAACAGGCUGAUUCUGA  9.9% 14863 CDS 222 1206ACCAGUUAAACAGGCUGAUUCUGGA  3.9% 14864 CDS 225 1207AGUUAAACAGGCUGAUUCUGGAAGA 20.48%  14865 CDS 226 1208GUUAAACAGGCUGAUUCUGGAAGUA 10.7% 14866 CDS 227 1209UUAAACAGGCUGAUUCUGGAAGUUA 22.75%  14867 CDS 228 1210UAAACAGGCUGAUUCUGGAAGUUCA 0.26% 14868 CDS 234 1211GGCUGAUUCUGGAAGUUCUGAGGAA 0.34% 14869 CDS 236 1212CUGAUUCUGGAAGUUCUGAGGAAAA  4.4% 14870 CDS 238 1213GAUUCUGGAAGUUCUGAGGAAAAGA  4.5% 14871 CDS 239 1214AUUCUGGAAGUUCUGAGGAAAAGCA  7.5% 14872 CDS 240 1215UUCUGGAAGUUCUGAGGAAAAGCAA 101.3%  14873 CDS 338 1216CCCCACAGACCCUUCCAAGUAAGUA 48.3% 14874 CDS 340 1217CCACAGACCCUUCCAAGUAAGUCCA 33.9% 14875 CDS 342 1218ACAGACCCUUCCAAGUAAGUCCAAA 16.1% 14876 CDS 343 1219CAGACCCUUCCAAGUAAGUCCAACA 38.7% 14877 CDS 345 1220GACCCUUCCAAGUAAGUCCAACGAA 54.2% 14878 CDS 348 1221CCUUCCAAGUAAGUCCAACGAAAGA 12.54%  14879 CDS 349 1222CUUCCAAGUAAGUCCAACGAAAGCA 32.44%  14880 CDS 351 1223UCCAAGUAAGUCCAACGAAAGCCAA 17.1% 14881 CDS 353 1224CAAGUAAGUCCAACGAAAGCCAUGA 32.94%  14882 CDS 358 1225AAGUCCAACGAAAGCCAUGACCACA 65.1% 14883 CDS 362 1226CCAACGAAAGCCAUGACCACAUGGA 76.9% 14884 CDS 363 1227CAACGAAAGCCAUGACCACAUGGAA 69.8% 14885 CDS 366 1228CGAAAGCCAUGACCACAUGGAUGAA 78.02%  14886 CDS 372 1229CCAUGACCACAUGGAUGAUAUGGAA 19.49%  14887 CDS 377 1230ACCACAUGGAUGAUAUGGAUGAUGA 20.43%  14888 CDS 393 1231GGAUGAUGAAGAUGAUGAUGACCAA 29.1% 14889 CDS 394 1232GAUGAUGAAGAUGAUGAUGACCAUA 24.5% 14890 CDS 396 1233UGAUGAAGAUGAUGAUGACCAUGUA 25.90%  14891 CDS 398 1234AUGAAGAUGAUGAUGACCAUGUGGA 20.5% 14892 CDS 399 1235UGAAGAUGAUGAUGACCAUGUGGAA  7.9% 14893 CDS 430 1236GACUCCAUUGACUCGAACGACUCUA 21.6% 14894 CDS 431 1237ACUCCAUUGACUCGAACGACUCUGA 13.5% 14895 CDS 432 1238CUCCAUUGACUCGAACGACUCUGAA 12.33%  14896 CDS 435 1239CAUUGACUCGAACGACUCUGAUGAA 42.5% 14897 CDS 440 1240ACUCGAACGACUCUGAUGAUGUAGA 22.54%  14898 CDS 441 1241CUCGAACGACUCUGAUGAUGUAGAA 17.4% 14899 CDS 442 1242UCGAACGACUCUGAUGAUGUAGAUA 11.2% 14900 CDS 443 1243CGAACGACUCUGAUGAUGUAGAUGA 20.7% 14901 CDS 445 1244AACGACUCUGAUGAUGUAGAUGACA 27.1% 14902 CDS 449 1245ACUCUGAUGAUGUAGAUGACACUGA 39.8% 14903 CDS 450 1246CUCUGAUGAUGUAGAUGACACUGAA  9.6% 14904 CDS 451 1247UCUGAUGAUGUAGAUGACACUGAUA 4.44% 14905 CDS 452 1248CUGAUGAUGUAGAUGACACUGAUGA  8.7% 14906 CDS 453 1249UGAUGAUGUAGAUGACACUGAUGAA 16.72%  14907 CDS 461 1250UAGAUGACACUGAUGAUUCUCACCA 42.9% 14908 CDS 462 1251AGAUGACACUGAUGAUUCUCACCAA 30.1% 14909 CDS 469 1252ACUGAUGAUUCUCACCAGUCUGAUA  9.1% 14910 CDS 470 1253CUGAUGAUUCUCACCAGUCUGAUGA 19.0% 14911 CDS 471 1254UGAUGAUUCUCACCAGUCUGAUGAA 42.1% 14912 CDS 472 1255GAUGAUUCUCACCAGUCUGAUGAGA 59.1% 14913 CDS 476 1256AUUCUCACCAGUCUGAUGAGUCUCA 38.2% 14914 CDS 479 1257CUCACCAGUCUGAUGAGUCUCACCA 34.1% 14915 CDS 480 1258UCACCAGUCUGAUGAGUCUCACCAA 48.45%  14916 CDS 483 1259CCAGUCUGAUGAGUCUCACCAUUCA  9.5% 14917 CDS 484 1260CAGUCUGAUGAGUCUCACCAUUCUA 21.5% 14918 CDS 485 1261AGUCUGAUGAGUCUCACCAUUCUGA 18.6% 14919 CDS 486 1262GUCUGAUGAGUCUCACCAUUCUGAA 20.2% 14920 CDS 487 1263UCUGAUGAGUCUCACCAUUCUGAUA 10.9% 14921 CDS 488 1264CUGAUGAGUCUCACCAUUCUGAUGA 18.9% 14922 CDS 489 1265UGAUGAGUCUCACCAUUCUGAUGAA 10.7% 14923 CDS 490 1266GAUGAGUCUCACCAUUCUGAUGAAA 28.15%  14924 CDS 493 1267GAGUCUCACCAUUCUGAUGAAUCUA 18.33%  14925 CDS 495 1268GUCUCACCAUUCUGAUGAAUCUGAA 7.61% 14926 CDS 496 1269UCUCACCAUUCUGAUGAAUCUGAUA 2.99% 14927 CDS 497 1270CUCACCAUUCUGAUGAAUCUGAUGA 7.44% 14928 CDS 498 1271UCACCAUUCUGAUGAAUCUGAUGAA  9.7% 14929 CDS 499 1272CACCAUUCUGAUGAAUCUGAUGAAA 16.96%  14930 CDS 501 1273CCAUUCUGAUGAAUCUGAUGAACUA 3.08% 14931 CDS 505 1274UCUGAUGAAUCUGAUGAACUGGUCA 13.24%  14932 CDS 510 1275UGAAUCUGAUGAACUGGUCACUGAA 3.16% 14933 CDS 550 1276CCAGCAACCGAAGUUUUCACUCCAA 14.02%  14934 CDS 554 1277CAACCGAAGUUUUCACUCCAGUUGA 3.10% 14935 CDS 555 1278AACCGAAGUUUUCACUCCAGUUGUA 5.27% 14936 CDS 572 1279CAGUUGUCCCCACAGUAGACACAUA 13.2% 14937 CDS 573 1280AGUUGUCCCCACAGUAGACACAUAA 27.01%  14938 CDS 574 1281GUUGUCCCCACAGUAGACACAUAUA 8.76% 14939 CDS 588 1282AGACACAUAUGAUGGCCGAGGUGAA 14.04%  14940 CDS 589 1283GACACAUAUGAUGGCCGAGGUGAUA 18.40%  14941 CDS 598 1284GAUGGCCGAGGUGAUAGUGUGGUUA 12.50%  14942 CDS 601 1285GGCCGAGGUGAUAGUGUGGUUUAUA 13.76%  14943 CDS 602 1286GCCGAGGUGAUAGUGUGGUUUAUGA 5.34% 14944 CDS 603 1287CCGAGGUGAUAGUGUGGUUUAUGGA 29.69%  14945 CDS 604 1288CGAGGUGAUAGUGUGGUUUAUGGAA 33.34%  14946 CDS 606 1289AGGUGAUAGUGUGGUUUAUGGACUA 17.50%  14947 CDS 608 1290GUGAUAGUGUGGUUUAUGGACUGAA 45.90%  14948 CDS 609 1291UGAUAGUGUGGUUUAUGGACUGAGA 22.0% 14949 CDS 610 1292GAUAGUGUGGUUUAUGGACUGAGGA 19.93%  14950 CDS 611 1293AUAGUGUGGUUUAUGGACUGAGGUA 17.34%  14951 CDS 615 1294UGUGGUUUAUGGACUGAGGUCAAAA 5.60% 14952 CDS 617 1295UGGUUUAUGGACUGAGGUCAAAAUA 25.74%  14953 CDS 618 1296GGUUUAUGGACUGAGGUCAAAAUCA 17.63%  14954 CDS 619 1297GUUUAUGGACUGAGGUCAAAAUCUA 3.45% 14955 CDS 621 1298UUAUGGACUGAGGUCAAAAUCUAAA 18.03%  14956 CDS 622 1299UAUGGACUGAGGUCAAAAUCUAAGA 20.98%  14957 CDS 623 1300AUGGACUGAGGUCAAAAUCUAAGAA 20.60%  14958 CDS 624 1301UGGACUGAGGUCAAAAUCUAAGAAA 26.73%  14959 CDS 625 1302GGACUGAGGUCAAAAUCUAAGAAGA 7.45% 14960 CDS 626 1303GACUGAGGUCAAAAUCUAAGAAGUA 14.1% 14961 CDS 629 1304UGAGGUCAAAAUCUAAGAAGUUUCA 8.61% 14962 CDS 630 1305GAGGUCAAAAUCUAAGAAGUUUCGA 19.07%  14963 CDS 631 1306AGGUCAAAAUCUAAGAAGUUUCGCA 6.08% 14964 CDS 632 1307GGUCAAAAUCUAAGAAGUUUCGCAA 19.82%  14965 CDS 636 1308AAAAUCUAAGAAGUUUCGCAGACCA 21.55%  14966 CDS 637 1309AAAUCUAAGAAGUUUCGCAGACCUA 30.20%  14967 CDS 638 1310AAUCUAAGAAGUUUCGCAGACCUGA 18.23%  14968 CDS 686 1311ACGAGGACAUCACCUCACACAUGGA 14.85%  14969 CDS 687 1312CGAGGACAUCACCUCACACAUGGAA 28.04%  14970 CDS 689 1313AGGACAUCACCUCACACAUGGAAAA 3.80% 14971 CDS 698 1314CCUCACACAUGGAAAGCGAGGAGUA 7.67% 14972 CDS 703 1315CACAUGGAAAGCGAGGAGUUGAAUA  5.8% 14973 CDS 704 1316ACAUGGAAAGCGAGGAGUUGAAUGA  5.3% 14974 CDS 705 1317CAUGGAAAGCGAGGAGUUGAAUGGA 24.47%  14975 CDS 718 1318GAGUUGAAUGGUGCAUACAAGGCCA 26.39%  14976 CDS 785 1319GCCGUGGGAAGGACAGUUAUGAAAA 7.60% 14977 CDS 786 1320CCGUGGGAAGGACAGUUAUGAAACA 8.75% 14978 CDS 788 1321GUGGGAAGGACAGUUAUGAAACGAA 8.34% 14979 CDS 790 1322GGGAAGGACAGUUAUGAAACGAGUA 5.38% 14980 CDS 792 1323GAAGGACAGUUAUGAAACGAGUCAA 11.45%  14981 CDS 794 1324AGGACAGUUAUGAAACGAGUCAGCA 11.78%  14982 CDS 795 1325GGACAGUUAUGAAACGAGUCAGCUA 10.69%  14983 CDS 797 1326ACAGUUAUGAAACGAGUCAGCUGGA 54.58%  14984 CDS 798 1327CAGUUAUGAAACGAGUCAGCUGGAA 33.9% 14985 CDS 846 1328CCACAAGCAGUCCAGAUUAUAUAAA 24.1% 14986 CDS 850 1329AAGCAGUCCAGAUUAUAUAAGCGGA 27.86%  14987 CDS 854 1330AGUCCAGAUUAUAUAAGCGGAAAGA 24.29%  14988 CDS 855 1331GUCCAGAUUAUAUAAGCGGAAAGCA 54.43%  14989 CDS 859 1332AGAUUAUAUAAGCGGAAAGCCAAUA 71.49%  14990 CDS 860 1333GAUUAUAUAAGCGGAAAGCCAAUGA 69.64%  14991 CDS 861 1334AUUAUAUAAGCGGAAAGCCAAUGAA 38.82%  14992 CDS 862 1335UUAUAUAAGCGGAAAGCCAAUGAUA 20.77%  14993 CDS 865 1336UAUAAGCGGAAAGCCAAUGAUGAGA 21.79%  14994 CDS 866 1337AUAAGCGGAAAGCCAAUGAUGAGAA 50.00%  14995 CDS 867 1338UAAGCGGAAAGCCAAUGAUGAGAGA 11.67%  14996 CDS 870 1339GCGGAAAGCCAAUGAUGAGAGCAAA 13.5% 14997 CDS 871 1340CGGAAAGCCAAUGAUGAGAGCAAUA 15.49%  14998 CDS 872 1341GGAAAGCCAAUGAUGAGAGCAAUGA 8.55% 14999 CDS 873 1342GAAAGCCAAUGAUGAGAGCAAUGAA 12.12%  15000 CDS 875 1343AAGCCAAUGAUGAGAGCAAUGAGCA 16.14%  15001 CDS 878 1344CCAAUGAUGAGAGCAAUGAGCAUUA 31.71%  15002 CDS 879 1345CAAUGAUGAGAGCAAUGAGCAUUCA 32.25%  15003 CDS 881 1346AUGAUGAGAGCAAUGAGCAUUCCGA 6.97% 15004 CDS 883 1347GAUGAGAGCAAUGAGCAUUCCGAUA 23.11%  15005 CDS 885 1348UGAGAGCAAUGAGCAUUCCGAUGUA 5.53% 15006 CDS 890 1349GCAAUGAGCAUUCCGAUGUGAUUGA 10.69%  15007 CDS 893 1350AUGAGCAUUCCGAUGUGAUUGAUAA 4.12% 15008 CDS 894 1351UGAGCAUUCCGAUGUGAUUGAUAGA 6.49% 15009 CDS 895 1352GAGCAUUCCGAUGUGAUUGAUAGUA 29.12%  15010 CDS 897 1353GCAUUCCGAUGUGAUUGAUAGUCAA 3.54% 15011 CDS 899 1354AUUCCGAUGUGAUUGAUAGUCAGGA 6.05% 15012 CDS 901 1355UCCGAUGUGAUUGAUAGUCAGGAAA 3.31% 15013 CDS 906 1356UGUGAUUGAUAGUCAGGAACUUUCA 12.71%  15014 CDS 907 1357GUGAUUGAUAGUCAGGAACUUUCCA 13.95%  15015 CDS 909 1358GAUUGAUAGUCAGGAACUUUCCAAA 4.03% 15016 CDS 912 1359UGAUAGUCAGGAACUUUCCAAAGUC 11.96%  15017 CDS 913 1360GAUAGUCAGGAACUUUCCAAAGUCA 14.01%  15018 CDS 914 1361AUAGUCAGGAACUUUCCAAAGUCAA 5.56% 15019 CDS 916 1362AGUCAGGAACUUUCCAAAGUCAGCA 13.92%  15020 CDS 917 1363GUCAGGAACUUUCCAAAGUCAGCCA 19.00%  15021 CDS 923 1364AACUUUCCAAAGUCAGCCGUGAAUA 17.56%  15022 CDS 925 1365CUUUCCAAAGUCAGCCGUGAAUUCA 19.58%  15023 CDS 926 1366UUUCCAAAGUCAGCCGUGAAUUCCA 6.54% 15024 CDS 935 1367UCAGCCGUGAAUUCCACAGCCAUGA 16.15%  15025 CDS 936 1368CAGCCGUGAAUUCCACAGCCAUGAA 20.62%  15026 CDS 937 1369AGCCGUGAAUUCCACAGCCAUGAAA 5.21% 15027 CDS 943 1370GAAUUCCACAGCCAUGAAUUUCACA 31.14%  15028 CDS 944 1371AAUUCCACAGCCAUGAAUUUCACAA 35.63%  15029 CDS 945 1372AUUCCACAGCCAUGAAUUUCACAGA 23.96%  15030 CDS 946 1373UUCCACAGCCAUGAAUUUCACAGCA 15.20%  15031 CDS 947 1374UCCACAGCCAUGAAUUUCACAGCCA 19.45%  15032 CDS 950 1375ACAGCCAUGAAUUUCACAGCCAUGA 25.74%  15033 CDS 952 1376AGCCAUGAAUUUCACAGCCAUGAAA 2.59% 15034 CDS 953 1377GCCAUGAAUUUCACAGCCAUGAAGA 6.00% 15035 CDS 954 1378CCAUGAAUUUCACAGCCAUGAAGAA 4.60% 15036 CDS 956 1379AUGAAUUUCACAGCCAUGAAGAUAA 9.20% 15037 CDS 957 1380UGAAUUUCACAGCCAUGAAGAUAUA 10.84%  15038 CDS 958 1381GAAUUUCACAGCCAUGAAGAUAUGA 40.20%  15039 CDS 959 1382AAUUUCACAGCCAUGAAGAUAUGCA 37.25%  15040 CDS 960 1383AUUUCACAGCCAUGAAGAUAUGCUA 8.21% 15041 CDS 961 1384UUUCACAGCCAUGAAGAUAUGCUGA 12.01%  15042 CDS 964 1385CACAGCCAUGAAGAUAUGCUGGUUA 12.25%  15043 CDS 983 1386UGGUUGUAGACCCCAAAAGUAAGGA 19.65%  15044 CDS 984 1387GGUUGUAGACCCCAAAAGUAAGGAA 28.19%  15045 CDS 985 1388GUUGUAGACCCCAAAAGUAAGGAAA 17.92%  15046 CDS 986 1389UUGUAGACCCCAAAAGUAAGGAAGA 7.94% 15047 CDS 987 1390UGUAGACCCCAAAAGUAAGGAAGAA 15.09%  15048 CDS 988 1391GUAGACCCCAAAAGUAAGGAAGAAA 20.01%  15049 CDS 989 1392UAGACCCCAAAAGUAAGGAAGAAGA 7.25% 15050 CDS 990 1393AGACCCCAAAAGUAAGGAAGAAGAA 12.42%  15051 CDS 995 1394CCAAAAGUAAGGAAGAAGAUAAACA 8.96% 15052 CDS 996 1395CAAAAGUAAGGAAGAAGAUAAACAA 6.85% 15053 CDS 997 1396AAAAGUAAGGAAGAAGAUAAACACA 14.15%  15054 CDS 998 1397AAAGUAAGGAAGAAGAUAAACACCA 12.32%  15055 CDS 999 1398AAGUAAGGAAGAAGAUAAACACCUA 8.83% 15056 CDS 1001 1399GUAAGGAAGAAGAUAAACACCUGAA 15.09%  15057 CDS 1002 1400UAAGGAAGAAGAUAAACACCUGAAA 4.91% 15058 CDS 1007 1401AAGAAGAUAAACACCUGAAAUUUCA 1.43% 15059 CDS 1008 1402AGAAGAUAAACACCUGAAAUUUCGA 3.51% 15060 CDS 1009 1403GAAGAUAAACACCUGAAAUUUCGUA 15.12%  15061 CDS 1010 1404AAGAUAAACACCUGAAAUUUCGUAA 28.56%  15062 CDS 1013 1405AUAAACACCUGAAAUUUCGUAUUUA 5.74% 15063 CDS 1015 1406AAACACCUGAAAUUUCGUAUUUCUA 13.01%  15064 CDS 1024 1407AAAUUUCGUAUUUCUCAUGAAUUAA 15.54%  15065 CDS 1030 1408CGUAUUUCUCAUGAAUUAGAUAGUA 9.47% 15066 CDS 1031 1409GUAUUUCUCAUGAAUUAGAUAGUGA 30.03%  15067 CDS 1032 1410UAUUUCUCAUGAAUUAGAUAGUGCA 5.31% 15068 CDS 1036 1411UCUCAUGAAUUAGAUAGUGCAUCUA 9.74% 15069 CDS 1037 1412CUCAUGAAUUAGAUAGUGCAUCUUA 10.78%  15070 CDS 1038 1413UCAUGAAUUAGAUAGUGCAUCUUCA 91.87%  15071 CDS 1039 1414CAUGAAUUAGAUAGUGCAUCUUCUA 93.82%  15072 CDS 1040 1415AUGAAUUAGAUAGUGCAUCUUCUGA 96.06%  15073 CDS 1041 1416UGAAUUAGAUAGUGCAUCUUCUGAA 94.91%  15074 CDS 1042 1417GAAUUAGAUAGUGCAUCUUCUGAGA 97.91%  15075 CDS 1043 1418AAUUAGAUAGUGCAUCUUCUGAGGA 93.76%  15076 CDS 1044 1419AUUAGAUAGUGCAUCUUCUGAGGUA 103.92%   15077 CDS 1045 1420UUAGAUAGUGCAUCUUCUGAGGUCA 95.85%  15078 CDS/3UTR 1052 1421GUGCAUCUUCUGAGGUCAAUUAAAA 93.83%  15079 CDS/3UTR 1053 1422UGCAUCUUCUGAGGUCAAUUAAAAA 90.69%  15080 CDS/3UTR 1054 1423GCAUCUUCUGAGGUCAAUUAAAAGA 101.49%   15081 CDS/3UTR 1055 1424CAUCUUCUGAGGUCAAUUAAAAGGA 110.27%   15082 CDS/3UTR 1056 1425AUCUUCUGAGGUCAAUUAAAAGGAA 99.36%  15083 CDS/3UTR 1057 1426UCUUCUGAGGUCAAUUAAAAGGAGA 95.31%  15084 CDS/3UTR 1058 1427CUUCUGAGGUCAAUUAAAAGGAGAA 15.55%  15085 3UTR 1081 1428AAAAAAUACAAUUUCUCACUUUGCA 3.59% 15086 3UTR 1083 1429AAAAUACAAUUUCUCACUUUGCAUU 3.46% 15087 3UTR 1086 1430AUACAAUUUCUCACUUUGCAUUUAG 2.37% 15088 3UTR 1087 1431UACAAUUUCUCACUUUGCAUUUAGU 3.54% 15089 3UTR 1088 1432ACAAUUUCUCACUUUGCAUUUAGUC 2.85% 15090 3UTR 1089 1433CAAUUUCUCACUUUGCAUUUAGUCA 2.35% 15091 3UTR 1093 1434UUCUCACUUUGCAUUUAGUCAAAAG 1.38% 15092 3UTR 1125 1435GCUUUAUAGCAAAAUGAAAGAGAAC 4.11% 15093 3UTR 1127 1436UUUAUAGCAAAAUGAAAGAGAACAU 3.91% 15094 3UTR 1128 1437UUAUAGCAAAAUGAAAGAGAACAUG 3.59% 15095 3UTR 1147 1438AACAUGAAAUGCUUCUUUCUCAGUU 1.80% 15096 3UTR 1148 1439ACAUGAAAUGCUUCUUUCUCAGUUU 2.17% 15097 3UTR 1150 1440AUGAAAUGCUUCUUUCUCAGUUUAU 2.93% 15098 3UTR 1153 1441AAAUGCUUCUUUCUCAGUUUAUUGG 2.18% 15099 3UTR 1154 1442AAUGCUUCUUUCUCAGUUUAUUGGU 3.92% 15100 3UTR 1156 1443UGCUUCUUUCUCAGUUUAUUGGUUG 4.08% 15101 3UTR 1157 1444GCUUCUUUCUCAGUUUAUUGGUUGA 1.74% 15102 3UTR 1158 1445CUUCUUUCUCAGUUUAUUGGUUGAA 4.74% 15103 3UTR 1159 1446UUCUUUCUCAGUUUAUUGGUUGAAU 2.65% 15104 3UTR 1168 1447AGUUUAUUGGUUGAAUGUGUAUCUA 2.57% 15105 3UTR 1178 1448UUGAAUGUGUAUCUAUUUGAGUCUG 3.76% 15106 3UTR 1179 1449UGAAUGUGUAUCUAUUUGAGUCUGG 2.91% 15107 3UTR 1183 1450UGUGUAUCUAUUUGAGUCUGGAAAU 0.62% 15108 3UTR 1184 1451GUGUAUCUAUUUGAGUCUGGAAAUA 2.45% 15109 3UTR 1186 1452GUAUCUAUUUGAGUCUGGAAAUAAC 2.18% 15110 3UTR 1191 1453UAUUUGAGUCUGGAAAUAACUAAUG 2.44% 15111 3UTR 1218 1454UUUGAUAAUUAGUUUAGUUUGUGGC 19.35%  15112 3UTR 1219 1455UUGAUAAUUAGUUUAGUUUGUGGCU 6.19% 15113 3UTR 1222 1456AUAAUUAGUUUAGUUUGUGGCUUCA 3.25% 15114 3UTR 1224 1457AAUUAGUUUAGUUUGUGGCUUCAUG 2.47% 15115 3UTR 1225 1458AUUAGUUUAGUUUGUGGCUUCAUGG 2.28% 15116 3UTR 1226 1459UUAGUUUAGUUUGUGGCUUCAUGGA 3.40% 15117 3UTR 1227 1460UAGUUUAGUUUGUGGCUUCAUGGAA 4.12% 15118 3UTR 1244 1461UCAUGGAAACUCCCUGUAAACUAAA 2.63% 15119 3UTR 1245 1462CAUGGAAACUCCCUGUAAACUAAAA 2.20% 15120 3UTR 1246 1463AUGGAAACUCCCUGUAAACUAAAAG 3.56% 15121 3UTR 1247 1464UGGAAACUCCCUGUAAACUAAAAGC 3.73% 15122 3UTR 1248 1465GGAAACUCCCUGUAAACUAAAAGCU 2.43% 15123 3UTR 1249 1466GAAACUCCCUGUAAACUAAAAGCUU 2.28% 15124 3UTR 1251 1467AACUCCCUGUAAACUAAAAGCUUCA 5.40% 15125 3UTR 1253 1468CUCCCUGUAAACUAAAAGCUUCAGG 8.21% 15126 3UTR 1286 1469UAUGUUCAUUCUAUAGAAGAAAUGC 3.17% 15127 3UTR 1294 1470UUCUAUAGAAGAAAUGCAAACUAUC 2.45% 15128 3UTR 1295 1471UCUAUAGAAGAAAUGCAAACUAUCA 3.97% 15129 3UTR 1296 1472CUAUAGAAGAAAUGCAAACUAUCAC 3.86% 15130 3UTR 1297 1473UAUAGAAGAAAUGCAAACUAUCACU 1.84% 15131 3UTR 1299 1474UAGAAGAAAUGCAAACUAUCACUGU 2.53% 15132 3UTR 1302 1475AAGAAAUGCAAACUAUCACUGUAUU 2.25% 15133 3UTR 1303 1476AGAAAUGCAAACUAUCACUGUAUUU 3.32% 15134 3UTR 1357 1477AUUUAUGUAGAAGCAAACAAAAUAC 1.86% 15135 3UTR 1465 1478UAUCUUUUUGUGGUGUGAAUAAAUC 3.40% 15136 3UTR 1466 1479AUCUUUUUGUGGUGUGAAUAAAUCU 3.49% 15137 3UTR 1467 1480UCUUUUUGUGGUGUGAAUAAAUCUU 3.03% 15138 3UTR 1468 1481CUUUUUGUGGUGUGAAUAAAUCUUU 3.62% 15139 3UTR 1496 1482CUUGAAUGUAAUAAGAAUUUGGUGG 61.48%  15140 3UTR 1497 1483UUGAAUGUAAUAAGAAUUUGGUGGU 71.54%  15141 3UTR 1504 1484UAAUAAGAAUUUGGUGGUGUCAAUU 58.54%  15142 3UTR 1511 1485AAUUUGGUGGUGUCAAUUGCUUAUU 56.93%  15143 3UTR 1512 1486AUUUGGUGGUGUCAAUUGCUUAUUU 81.22%  15144 3UTR 1513 1487UUUGGUGGUGUCAAUUGCUUAUUUG 59.16%  15145 3UTR 1514 1488UUGGUGGUGUCAAUUGCUUAUUUGU 59.46%  15146 3UTR 1540 1489UUCCCACGGUUGUCCAGCAAUUAAU 67.40% 

TABLE 10 hCTGF sd-rxRNA CTGF Target Gene Single Strand SEQ ID Duplex IDID sd-rxRNA sequence NO 17356 17007 A.mC.A.mU.mU.A.A.mC.mU.mC.A.mU.A.Chl1490 17009 P.mU.A.fU.G.A.G.fU.fU.A.A.fU.G.fU*fC*fU*fC*fU*fC*A 1491 1735717008 G.A.mC.A.mU.mU.A.A.mC.mU.mC.A.mU.A.Chl 1492 17009P.mU.A.fU.G.A.G.fU.fU.A.A.fU.G.fU*fC*fU*fC*fU*fC*A 1493 17358 17010mU.G.A.A.G.A.A.mU.G.mU.mU.A.A.Chl 1494 17012P.mU.fU.A.A.fC.A.fU.fU.fC.fU.fU.fC.A*A*A*fC*fC*A*G 1495 17359 17011mU.mU.G.A.A.G.A.A.mU.G.mU.mU.A.A.Chl 1496 17012P.mU.fU.A.A.fC.A.fU.fU.fC.fU.fU.fC.A*A*A*fC*fC*A*G 1497 17360 17013G.A.mU.A.G.mC.A.mU.mC.mU.mU.A.A.Chl 1498 17015P.mU.fU.A.A.G.A.fU.G.fC.fU.A.fU.fC*fU*G*A*fU*G*A 1499 17361 17014A.G.A.mU.A.G.mC.A.mU.mC.mU.mU.A.A.Chl 1500 17015P.mU.fU.A.A.G.A.fU.G.fC.fU.A.fU.fC*fU*G*A*fU*G*A 1501 17362 17016mU.G.A.A.G.mU.G.mU.A.A.mU.mU.A.Chl 1502 17017P.mU.A.A.fU.fU.A.fC.A.fC.fU.fU.fC.A*A*A*fU*A*G*C 1503 17363 17018A.A.mU.mU.G.A.G.A.A.G.G.A.A.Chl 1504 17019P.mU.fU.fC.fC.fU.fU.fC.fU.fC.A.A.fU.fU*A*fC*A*fC*fU*U 1505 17364 17020mU.mU.G.A.G.A.A.G.G.A.A.A.A.Chl 1506 17021P.mU.fU.fU.fU.fC.fC.fU.fU.fC.fU.fC.A.A*fU*fU*A*fC*A*C 1507 17365 17022mC.A.mU.mU.mC.mU.G.A.mU.mU.mC.G.A.Chl 1508 17023P.mU.fC.G.A.A.fU.fC.A.G.A.A.fU.G*fU*fC*A*G*A*G 1509 17366 17024mU.mU.mC.mU.G.A.mU.mU.mC.G.A.A.A.Chl 1510 17025P.mU.fU.fU.fC.G.A.A.fU.fC.A.G.A.A*fU*G*fU*fC*A*G 1511 17367 17026mC.mU.G.mU.mC.G.A.mU.mU.A.G.A.A.Chl 1512 17027P.mU.fU.fC.fU.A.A.fU.fC.G.A.fC.A.G*G*A*fU*fU*fC*C 1513 17368 17028mU.mU.mU.G.mC.mC.mU.G.mU.A.A.mC.A.Chl 1514 17030P.mU.G.fU.fU.A.fC.A.G.G.fC.A.A.A*fU*fU*fC*A*fC*U 1515 17369 17029A.mU.mU.mU.G.mC.mC.mU.G.mU.A.A.mC.A.Chl 1516 17030P.mU.G.fU.fU.A.fC.A.G.G.fC.A.A.A*fU*fU*fC*A*fC*U 1517 17370 17031A.mC.A.A.G.mC.mC.A.G.A.mU.mU.A.Chl 1518 17033P.mU.A.A.fU.fC.fU.G.G.fC.fU.fU.G.fU*fU*A*fC*A*G*G 1519 17371 17032“A.A.mC.A.A.G.mC.mC.A.G.A.mU.mU.A.Chl 1520 17033P.mU.A.A.fU.fC.fU.G.G.fC.fU.fU.G.fU*fU*A*fC*A*G*G 1521 17372 17034mC.A.G.mU.mU.mU.A.mU.mU.mU.G.mU.A.Chl 1522 17035P.mU.A.fC.A.A.A.fU.A.A.A.fC.fU.G*fU*fC*fC*G*A*A 1523 17373 17036mU.G.mU.mU.G.A.G.A.G.mU.G.mU.A.Chl 1524 17038P.mU.A.fC.A.fC.fU.fC.fU.fC.A.A.fC.A*A*A*fU*A*A*A 1525 17374 17037mU.mU.G.mU.mU.G.A.G.A.G.mU.G.mU.A.Chl 1526 17038P.mU.A.fC.A.fC.fU.fC.fU.fC.A.A.fC.A*A*A*fU*A*A*A 1527 17375 17039mU.G.mC.A.mC.mC.mU.mU.mU.mC.mU.A.A.Chl 1528 17041P.mU.fU.A.G.A.A.A.G.G.fU.G.fC.A*A*A*fC*A*fU*G 1529 17376 17040mU.mU.G.mC.A.mC.mC.mU.mU.mU.mC.mU.A.A.Chl 1530 17041P.mU.fU.A.G.A.A.A.G.G.fU.G.fC.A*A*A*fC*A*fU*G 1531 17377 17042mU.mU.G.A.G.mC.mU.mU.mU.mC.mU.G.A.Chl 1532 17043P.mU.fC.A.G.A.A.A.G.fC.fU.fC.A.A*A*fC*fU*fU*G*A 1533 17378 17044mU.G.A.G.A.G.mU.G.mU.G.A.mC.A.Chl 1534 17045P.mU.G.fU.fC.A.fC.A.fC.fU.fC.fU.fC.A*A*fC*A*A*A*U 1535 17379 17046A.G.mU.G.mU.G.A.mC.mC.A.A.A.A.Chl 1536 17048P.mU.fU.fU.fU.G.G.fU.fC.A.fC.A.fC.fU*fC*fU*fC*A*A*C 1537 17380 17047G.A.G.mU.G.mU.G.A.mC.mC.A.A.A.A.Chl 1538 17048P.mU.fU.fU.fU.G.G.fU.fC.A.fC.A.fC.fU*fC*fU*fC*A*A*C 1539 17381 17049G.mU.G.mU.G.A.mC.mC.A.A.A.A.A.Chl 1540 17050P.mU.fU.fU.fU.fU.G.G.fU.fC.A.fC.A.fC*fU*fC*fU*fC*A*A 1541 17382 17051mU.G.mU.G.A.mC.mC.A.A.A.A.G.A.Chl 1542 17053P.mU.fC.fU.fU.fU.fU.G.G.fU.fC.A.fC.A*fC*fU*fC*fU*fC*A 1543 17383 17052G.mU.G.mU.G.A.mC.mC.A.A.A.A.G.A.Chl 1544 17053P.mU.fC.fU.fU.fU.fU.G.G.fU.fC.A.fC.A*fC*fU*fC*fU*fC*A 1545 17384 17054G.mU.G.A.mC.mC.A.A.A.A.G.mU.A.Chl 1546 17055P.mU.A.fC.fU.fU.fU.fU.G.G.fU.fC.A.fC*A*fC*fU*fC*fU*C 1547 17385 17056G.A.mC.mC.A.A.A.A.G.mU.mU.A.A.Chl 1548 17057P.mU.fU.A.A.fC.fU.fU.fU.fU.G.G.fU.fC*A*fC*A*fC*fU*C 1549 17386 17058G.mC.A.mC.mC.mU.mU.mU.mC.mU.A.G.A.Chl 1550 17059P.mU.fC.fU.A.G.A.A.A.G.G.fU.G.fC*A*A*A*fC*A*U 1551 17387 17060mC.mC.mU.mU.mU.mC.mU.A.G.mU.mU.G.A.Chl 1552 17061P.mU.fC.A.A.fC.fU.A.G.A.A.A.G.G*fU*G*fC*A*A*A 1553

TABLE 11 Inhibition of gene expression with hCTGF ori sequences TargetGene Gene Ref SEQ ID CTGF % Expression Duplex Name Region Pos NO Sensesequence (0.1 nM) 14542 CDS 774 1554 UUUGGCCCAGACCCAACUAUGAUUA 96% 14543CDS 776 1555 UGGCCCAGACCCAACUAUGAUUAGA 94% 14544 CDS 785 1556CCCAACUAUGAUUAGAGCCAACUGA 55% 14545 CDS 786 1557CCAACUAUGAUUAGAGCCAACUGCA 89% 14546 CDS 934 1558CUUGCGAAGCUGACCUGGAAGAGAA 63% 14547 CDS 938 1559CGAAGCUGACCUGGAAGAGAACAUA 70% 14548 CDS 940 1560AAGCUGACCUGGAAGAGAACAUUAA 65% 14549 CDS 941 1561AGCUGACCUGGAAGAGAACAUUAAA 81% 14550 CDS 943 1562CUGACCUGGAAGAGAACAUUAAGAA 85% 14551 CDS 944 1563UGACCUGGAAGAGAACAUUAAGAAA 61% 14552 CDS 945 1564GACCUGGAAGAGAACAUUAAGAAGA 73% 14553 CDS 983 1565CCGUACUCCCAAAAUCUCCAAGCCA 86% 14554 CDS 984 1566CGUACUCCCAAAAUCUCCAAGCCUA 64% 14555 CDS 985 1567GUACUCCCAAAAUCUCCAAGCCUAA 71% 14556 CDS 986 1568UACUCCCAAAAUCUCCAAGCCUAUA 71% 14557 CDS 987 1569ACUCCCAAAAUCUCCAAGCCUAUCA 84% 14558 CDS 988 1570CUCCCAAAAUCUCCAAGCCUAUCAA 64% 14559 CDS 989 1571UCCCAAAAUCUCCAAGCCUAUCAAA 64% 14560 CDS 990 1572CCCAAAAUCUCCAAGCCUAUCAAGA 87% 14561 CDS 1002 1573AAGCCUAUCAAGUUUGAGCUUUCUA 46% 14562 CDS 1003 1574AGCCUAUCAAGUUUGAGCUUUCUGA 30% 14563 CDS 1004 1575GCCUAUCAAGUUUGAGCUUUCUGGA 63% 14564 CDS 1008 1576AUCAAGUUUGAGCUUUCUGGCUGCA 77% 14565 CDS 1025 1577UGGCUGCACCAGCAUGAAGACAUAA 96% 14566 CDS 1028 1578CUGCACCAGCAUGAAGACAUACCGA 79% 14567 CDS 1029 1579UGCACCAGCAUGAAGACAUACCGAA 58% 14568 CDS 1033 1580CCAGCAUGAAGACAUACCGAGCUAA 59% 14569 CDS 1035 1581AGCAUGAAGACAUACCGAGCUAAAA 76% 14570 CDS 1036 1582GCAUGAAGACAUACCGAGCUAAAUA 71% 14571 CDS 1050 1583CGAGCUAAAUUCUGUGGAGUAUGUA 73% 14572 CDS 1051 1584GAGCUAAAUUCUGUGGAGUAUGUAA 72% 14573 CDS 1053 1585GCUAAAUUCUGUGGAGUAUGUACCA 87% 14574 CDS 1054 1586CUAAAUUCUGUGGAGUAUGUACCGA 83% 14575 CDS 1135 1587CUGACGGCGAGGUCAUGAAGAAGAA 77% 14576 CDS 1138 1588ACGGCGAGGUCAUGAAGAAGAACAA 72% 14577 CDS 1139 1589CGGCGAGGUCAUGAAGAAGAACAUA 85% 14578 CDS 1143 1590GAGGUCAUGAAGAAGAACAUGAUGA 83% 14579 CDS 1145 1591GGUCAUGAAGAAGAACAUGAUGUUA 91% 14580 CDS 1148 1592CAUGAAGAAGAACAUGAUGUUCAUA 92% 14581 CDS 1157 1593GAACAUGAUGUUCAUCAAGACCUGA 84% 14582 CDS 1161 1594AUGAUGUUCAUCAAGACCUGUGCCA 92% 14583 CDS 1203 1595GGAGACAAUGACAUCUUUGAAUCGA 62% 14584 CDS 1204 1596GAGACAAUGACAUCUUUGAAUCGCA 56% 14585 CDS 1205 1597AGACAAUGACAUCUUUGAAUCGCUA 30% 14586 CDS 1206 1598GACAAUGACAUCUUUGAAUCGCUGA 47% 14587 CDS 1207 1599ACAAUGACAUCUUUGAAUCGCUGUA 29% 14588 CDS 1208 1600CAAUGACAUCUUUGAAUCGCUGUAA 50% 14589 CDS 1209 1601AAUGACAUCUUUGAAUCGCUGUACA 39% 14590 CDS 1210 1602AUGACAUCUUUGAAUCGCUGUACUA 44% 14591 CDS 1211 1603UGACAUCUUUGAAUCGCUGUACUAA 39% 14592 CDS 1212 1604GACAUCUUUGAAUCGCUGUACUACA 55% 14593 CDS 1213 1605ACAUCUUUGAAUCGCUGUACUACAA 59% 14594 CDS 1216 1606UCUUUGAAUCGCUGUACUACAGGAA 80% 14595 CDS 1217 1607CUUUGAAUCGCUGUACUACAGGAAA 80% 14596 CDS 1223 1608AUCGCUGUACUACAGGAAGAUGUAA 59% 14597 CDS 1224 1609UCGCUGUACUACAGGAAGAUGUACA 62% 14598 CDS 1239 1610AAGAUGUACGGAGACAUGGCAUGAA 59% 14599 CDS 1253 1611CAUGGCAUGAAGCCAGAGAGUGAGA 65% 14600 3UTR 1266 1612CAGAGAGUGAGAGACAUUAACUCAA 43% 14601 3UTR 1267 1613AGAGAGUGAGAGACAUUAACUCAUA 25% 14602 3UTR 1268 1614GAGAGUGAGAGACAUUAACUCAUUA 33% 14603 3UTR 1269 1615AGAGUGAGAGACAUUAACUCAUUAA 42% 14604 3UTR 1270 1616GAGUGAGAGACAUUAACUCAUUAGA 28% 14605 3UTR 1271 1617AGUGAGAGACAUUAACUCAUUAGAA 34% 14606 3UTR 1272 1618GUGAGAGACAUUAACUCAUUAGACA 30% 14607 3UTR 1273 1619UGAGAGACAUUAACUCAUUAGACUA 33% 14608 3UTR 1275 1620AGAGACAUUAACUCAUUAGACUGGA 42% 14609 3UTR 1277 1621AGACAUUAACUCAUUAGACUGGAAA 25% 14610 3UTR 1278 1622GACAUUAACUCAUUAGACUGGAACA 31% 14611 3UTR 1279 1623ACAUUAACUCAUUAGACUGGAACUA 32% 14612 3UTR 1281 1624AUUAACUCAUUAGACUGGAACUUGA 23% 14613 3UTR 1284 1625AACUCAUUAGACUGGAACUUGAACA 39% 14614 3UTR 1285 1626ACUCAUUAGACUGGAACUUGAACUA 30% 14615 3UTR 1286 1627CUCAUUAGACUGGAACUUGAACUGA 43% 14616 3UTR 1287 1628UCAUUAGACUGGAACUUGAACUGAA 26% 14617 3UTR 1291 1629UAGACUGGAACUUGAACUGAUUCAA 33% 14618 3UTR 1293 1630GACUGGAACUUGAACUGAUUCACAA 43% 14619 3UTR 1294 1631ACUGGAACUUGAACUGAUUCACAUA 28% 14620 3UTR 1295 1632CUGGAACUUGAACUGAUUCACAUCA 41% 14621 3UTR 1296 1633UGGAACUUGAACUGAUUCACAUCUA 34% 14622 3UTR 1298 1634GAACUUGAACUGAUUCACAUCUCAA 31% 14623 3UTR 1299 1635AACUUGAACUGAUUCACAUCUCAUA 31% 14624 3UTR 1300 1636ACUUGAACUGAUUCACAUCUCAUUA 33% 14625 3UTR 1301 1637CUUGAACUGAUUCACAUCUCAUUUA 28% 14626 3UTR 1326 1638UCCGUAAAAAUGAUUUCAGUAGCAA 30% 14627 3UTR 1332 1639AAAAUGAUUUCAGUAGCACAAGUUA 28% 14628 3UTR 1395 1640CCCAAUUCAAAACAUUGUGCCAUGA 63% 14629 3UTR 1397 1641CAAUUCAAAACAUUGUGCCAUGUCA 39% 14630 3UTR 1402 1642CAAAACAUUGUGCCAUGUCAAACAA 34% 14631 3UTR 1408 1643AUUGUGCCAUGUCAAACAAAUAGUA 33% 14632 3UTR 1409 1644UUGUGCCAUGUCAAACAAAUAGUCA 33% 14633 3UTR 1412 1645UGCCAUGUCAAACAAAUAGUCUAUA 36% 14634 3UTR 1416 1646AUGUCAAACAAAUAGUCUAUCAACA 30% 14635 3UTR 1435 1647UCAACCCCAGACACUGGUUUGAAGA 39% 14636 3UTR 1436 1648CAACCCCAGACACUGGUUUGAAGAA 47% 14637 3UTR 1438 1649ACCCCAGACACUGGUUUGAAGAAUA 45% 14638 3UTR 1439 1650CCCCAGACACUGGUUUGAAGAAUGA 40% 14639 3UTR 1442 1651CAGACACUGGUUUGAAGAAUGUUAA 21% 14640 3UTR 1449 1652UGGUUUGAAGAAUGUUAAGACUUGA 22% 14641 3UTR 1453 1653UUGAAGAAUGUUAAGACUUGACAGA 24% 14642 3UTR 1454 1654UGAAGAAUGUUAAGACUUGACAGUA 37% 14643 3UTR 1462 1655GUUAAGACUUGACAGUGGAACUACA 20% 14644 3UTR 1470 1656UUGACAGUGGAACUACAUUAGUACA 30% 14645 3UTR 1471 1657UGACAGUGGAACUACAUUAGUACAA 43% 14646 3UTR 1474 1658CAGUGGAACUACAUUAGUACACAGA 36% 14647 3UTR 1475 1659AGUGGAACUACAUUAGUACACAGCA 38% 14648 3UTR 1476 1660GUGGAACUACAUUAGUACACAGCAA 35% 14649 3UTR 1477 1661UGGAACUACAUUAGUACACAGCACA 34% 14650 3UTR 1478 1662GGAACUACAUUAGUACACAGCACCA 33% 14651 3UTR 1479 1663GAACUACAUUAGUACACAGCACCAA 39% 14652 3UTR 1480 1664AACUACAUUAGUACACAGCACCAGA 27% 14653 3UTR 1481 1665ACUACAUUAGUACACAGCACCAGAA 29% 14654 3UTR 1482 1666CUACAUUAGUACACAGCACCAGAAA 38% 14655 3UTR 1483 1667UACAUUAGUACACAGCACCAGAAUA 28% 14656 3UTR 1484 1668ACAUUAGUACACAGCACCAGAAUGA 31% 14657 3UTR 1486 1669AUUAGUACACAGCACCAGAAUGUAA 26% 14658 3UTR 1487 1670UUAGUACACAGCACCAGAAUGUAUA 31% 14659 3UTR 1489 1671AGUACACAGCACCAGAAUGUAUAUA 35% 14660 3UTR 1490 1672GUACACAGCACCAGAAUGUAUAUUA 34% 14661 3UTR 1497 1673GCACCAGAAUGUAUAUUAAGGUGUA 32% 14662 3UTR 1503 1674GAAUGUAUAUUAAGGUGUGGCUUUA 42% 14663 3UTR 1539 1675AGGGUACCAGCAGAAAGGUUAGUAA 28% 14664 3UTR 1543 1676UACCAGCAGAAAGGUUAGUAUCAUA 29% 14665 3UTR 1544 1677ACCAGCAGAAAGGUUAGUAUCAUCA 33% 14666 3UTR 1548 1678GCAGAAAGGUUAGUAUCAUCAGAUA 34% 14667 3UTR 1557 1679UUAGUAUCAUCAGAUAGCAUCUUAA 22% 14668 3UTR 1576 1680UCUUAUACGAGUAAUAUGCCUGCUA 48% 14669 3UTR 1577 1681CUUAUACGAGUAAUAUGCCUGCUAA 31% 14670 3UTR 1579 1682UAUACGAGUAAUAUGCCUGCUAUUA 43% 14671 3UTR 1580 1683AUACGAGUAAUAUGCCUGCUAUUUA 39% 14672 3UTR 1581 1684UACGAGUAAUAUGCCUGCUAUUUGA 33% 14673 3UTR 1582 1685ACGAGUAAUAUGCCUGCUAUUUGAA 40% 14674 3UTR 1584 1686GAGUAAUAUGCCUGCUAUUUGAAGA 38% 14675 3UTR 1585 1687AGUAAUAUGCCUGCUAUUUGAAGUA 24% 14676 3UTR 1586 1688GUAAUAUGCCUGCUAUUUGAAGUGA 34% 14677 3UTR 1587 1689UAAUAUGCCUGCUAUUUGAAGUGUA 26% 14678 3UTR 1589 1690AUAUGCCUGCUAUUUGAAGUGUAAA 26% 14679 3UTR 1591 1691AUGCCUGCUAUUUGAAGUGUAAUUA 25% 14680 3UTR 1596 1692UGCUAUUUGAAGUGUAAUUGAGAAA 36% 14681 3UTR 1599 1693UAUUUGAAGUGUAAUUGAGAAGGAA 22% 14682 3UTR 1600 1694AUUUGAAGUGUAAUUGAGAAGGAAA 22% 14683 3UTR 1601 1695UUUGAAGUGUAAUUGAGAAGGAAAA 19% 14684 3UTR 1609 1696GUAAUUGAGAAGGAAAAUUUUAGCA 53% 14685 3UTR 1610 1697UAAUUGAGAAGGAAAAUUUUAGCGA 55% 14686 3UTR 1611 1698AAUUGAGAAGGAAAAUUUUAGCGUA 20% 14687 3UTR 1612 1699AUUGAGAAGGAAAAUUUUAGCGUGA 23% 14688 3UTR 1613 1700UUGAGAAGGAAAAUUUUAGCGUGCA 37% 14689 3UTR 1614 1701UGAGAAGGAAAAUUUUAGCGUGCUA 31% 14690 3UTR 1619 1702AGGAAAAUUUUAGCGUGCUCACUGA 46% 14691 3UTR 1657 1703CCAGUGACAGCUAGGAUGUGCAUUA 42% 14692 3UTR 1661 1704UGACAGCUAGGAUGUGCAUUCUCCA 39% 14693 3UTR 1682 1705UCCAGCCAUCAAGAGACUGAGUCAA 53% 14694 3UTR 1685 1706AGCCAUCAAGAGACUGAGUCAAGUA 71% 14695 3UTR 1686 1707GCCAUCAAGAGACUGAGUCAAGUUA 54% 14696 3UTR 1687 1708CCAUCAAGAGACUGAGUCAAGUUGA 71% 14697 3UTR 1688 1709CAUCAAGAGACUGAGUCAAGUUGUA 74% 14698 3UTR 1689 1710AUCAAGAGACUGAGUCAAGUUGUUA 61% 14699 3UTR 1690 1711UCAAGAGACUGAGUCAAGUUGUUCA 59% 14700 3UTR 1691 1712CAAGAGACUGAGUCAAGUUGUUCCA 73% 14701 3UTR 1692 1713AAGAGACUGAGUCAAGUUGUUCCUA 78% 14702 3UTR 1693 1714AGAGACUGAGUCAAGUUGUUCCUUA 60% 14703 3UTR 1695 1715AGACUGAGUCAAGUUGUUCCUUAAA 63% 14704 3UTR 1696 1716GACUGAGUCAAGUUGUUCCUUAAGA 92% 14705 3UTR 1697 1717ACUGAGUCAAGUUGUUCCUUAAGUA 74% 14706 3UTR 1707 1718GUUGUUCCUUAAGUCAGAACAGCAA 70% 14707 3UTR 1724 1719AACAGCAGACUCAGCUCUGACAUUA 69% 14708 3UTR 1725 1720ACAGCAGACUCAGCUCUGACAUUCA 67% 14709 3UTR 1726 1721CAGCAGACUCAGCUCUGACAUUCUA 71% 14710 3UTR 1727 1722AGCAGACUCAGCUCUGACAUUCUGA 73% 14711 3UTR 1728 1723GCAGACUCAGCUCUGACAUUCUGAA 60% 14712 3UTR 1729 1724CAGACUCAGCUCUGACAUUCUGAUA 72% 14713 3UTR 1732 1725ACUCAGCUCUGACAUUCUGAUUCGA 24% 14714 3UTR 1733 1726CUCAGCUCUGACAUUCUGAUUCGAA 32% 14715 3UTR 1734 1727UCAGCUCUGACAUUCUGAUUCGAAA 23% 14716 3UTR 1735 1728CAGCUCUGACAUUCUGAUUCGAAUA 27% 14717 3UTR 1736 1729AGCUCUGACAUUCUGAUUCGAAUGA 38% 14718 3UTR 1739 1730UCUGACAUUCUGAUUCGAAUGACAA 28% 14719 3UTR 1741 1731UGACAUUCUGAUUCGAAUGACACUA 29% 14720 3UTR 1742 1732GACAUUCUGAUUCGAAUGACACUGA 33% 14721 3UTR 1743 1733ACAUUCUGAUUCGAAUGACACUGUA 28% 14722 3UTR 1747 1734UCUGAUUCGAAUGACACUGUUCAGA 39% 14723 3UTR 1748 1735CUGAUUCGAAUGACACUGUUCAGGA 36% 14724 3UTR 1750 1736GAUUCGAAUGACACUGUUCAGGAAA 33% 14725 3UTR 1751 1737AUUCGAAUGACACUGUUCAGGAAUA 30% 14726 3UTR 1759 1738GACACUGUUCAGGAAUCGGAAUCCA 34% 14727 3UTR 1760 1739ACACUGUUCAGGAAUCGGAAUCCUA 35% 14728 3UTR 1761 1740CACUGUUCAGGAAUCGGAAUCCUGA 40% 14729 3UTR 1768 1741CAGGAAUCGGAAUCCUGUCGAUUAA 34% 14730 3UTR 1769 1742AGGAAUCGGAAUCCUGUCGAUUAGA 31% 14731 3UTR 1770 1743GGAAUCGGAAUCCUGUCGAUUAGAA 24% 14732 3UTR 1771 1744GAAUCGGAAUCCUGUCGAUUAGACA 32% 14733 3UTR 1772 1745AAUCGGAAUCCUGUCGAUUAGACUA 29% 14734 3UTR 1774 1746UCGGAAUCCUGUCGAUUAGACUGGA 34% 14735 3UTR 1777 1747GAAUCCUGUCGAUUAGACUGGACAA 51% 14736 3UTR 1782 1748CUGUCGAUUAGACUGGACAGCUUGA 88% 14737 3UTR 1783 1749UGUCGAUUAGACUGGACAGCUUGUA 38% 14738 3UTR 1797 1750GACAGCUUGUGGCAAGUGAAUUUGA 46% 14739 3UTR 1798 1751ACAGCUUGUGGCAAGUGAAUUUGCA 52% 14740 3UTR 1800 1752AGCUUGUGGCAAGUGAAUUUGCCUA 43% 14741 3UTR 1801 1753GCUUGUGGCAAGUGAAUUUGCCUGA 51% 14742 3UTR 1802 1754CUUGUGGCAAGUGAAUUUGCCUGUA 32% 14743 3UTR 1803 1755UUGUGGCAAGUGAAUUUGCCUGUAA 31% 14744 3UTR 1804 1756UGUGGCAAGUGAAUUUGCCUGUAAA 29% 14745 3UTR 1805 1757GUGGCAAGUGAAUUUGCCUGUAACA 20% 14746 3UTR 1806 1758UGGCAAGUGAAUUUGCCUGUAACAA 34% 14747 3UTR 1807 1759GGCAAGUGAAUUUGCCUGUAACAAA 31% 14748 3UTR 1808 1760GCAAGUGAAUUUGCCUGUAACAAGA 27% 14749 3UTR 1809 1761CAAGUGAAUUUGCCUGUAACAAGCA 34% 14750 3UTR 1810 1762AAGUGAAUUUGCCUGUAACAAGCCA 36% 14751 3UTR 1811 1763AGUGAAUUUGCCUGUAACAAGCCAA 31% 14752 3UTR 1814 1764GAAUUUGCCUGUAACAAGCCAGAUA 24% 14753 3UTR 1815 1765AAUUUGCCUGUAACAAGCCAGAUUA 21% 14754 3UTR 1816 1766AUUUGCCUGUAACAAGCCAGAUUUA 22% 14755 3UTR 1910 1767AAGUUAAUUUAAAGUUGUUUGUGCA 58% 14756 3UTR 1911 1768AGUUAAUUUAAAGUUGUUUGUGCCA 73% 14757 3UTR 1912 1769GUUAAUUUAAAGUUGUUUGUGCCUA 64% 14758 3UTR 1957 1770UUUGAUAUUUCAAUGUUAGCCUCAA 42% 14759 3UTR 1961 1771AUAUUUCAAUGUUAGCCUCAAUUUA 30% 14760 3UTR 1971 1772GUUAGCCUCAAUUUCUGAACACCAA 34% 14761 3UTR 1974 1773AGCCUCAAUUUCUGAACACCAUAGA 35% 14762 3UTR 1975 1774GCCUCAAUUUCUGAACACCAUAGGA 33% 14763 3UTR 1976 1775CCUCAAUUUCUGAACACCAUAGGUA 39% 14764 3UTR 1977 1776CUCAAUUUCUGAACACCAUAGGUAA 27% 14765 3UTR 1978 1777UCAAUUUCUGAACACCAUAGGUAGA 31% 14766 3UTR 1979 1778CAAUUUCUGAACACCAUAGGUAGAA 49% 14767 3UTR 1980 1779AAUUUCUGAACACCAUAGGUAGAAA 46% 14768 3UTR 1981 1780AUUUCUGAACACCAUAGGUAGAAUA 40% 14769 3UTR 1982 1781UUUCUGAACACCAUAGGUAGAAUGA 47% 14770 3UTR 1985 1782CUGAACACCAUAGGUAGAAUGUAAA 33% 14771 3UTR 1986 1783UGAACACCAUAGGUAGAAUGUAAAA 35% 14772 3UTR 1987 1784GAACACCAUAGGUAGAAUGUAAAGA 31% 14773 3UTR 1988 1785AACACCAUAGGUAGAAUGUAAAGCA 30% 14774 3UTR 1989 1786ACACCAUAGGUAGAAUGUAAAGCUA 32% 14775 3UTR 1991 1787ACCAUAGGUAGAAUGUAAAGCUUGA 31% 14776 3UTR 1992 1788CCAUAGGUAGAAUGUAAAGCUUGUA 34% 14777 3UTR 1993 1789CAUAGGUAGAAUGUAAAGCUUGUCA 31% 14778 3UTR 1994 1790AUAGGUAGAAUGUAAAGCUUGUCUA 28% 14779 3UTR 1996 1791AGGUAGAAUGUAAAGCUUGUCUGAA 32% 14780 3UTR 2002 1792AAUGUAAAGCUUGUCUGAUCGUUCA 34% 14781 3UTR 2017 1793UGAUCGUUCAAAGCAUGAAAUGGAA 31% 14782 3UTR 2021 1794CGUUCAAAGCAUGAAAUGGAUACUA 39% 14783 3UTR 2022 1795GUUCAAAGCAUGAAAUGGAUACUUA 25% 14784 3UTR 2023 1796UUCAAAGCAUGAAAUGGAUACUUAA 22% 14785 3UTR 2047 1797UAUGGAAAUUCUGCUCAGAUAGAAA 39% 14786 3UTR 2048 1798AUGGAAAUUCUGCUCAGAUAGAAUA 35% 14787 3UTR 2059 1799GCUCAGAUAGAAUGACAGUCCGUCA 44% 14788 3UTR 2060 1800CUCAGAUAGAAUGACAGUCCGUCAA 41% 14789 3UTR 2062 1801CAGAUAGAAUGACAGUCCGUCAAAA 46% 14790 3UTR 2063 1802AGAUAGAAUGACAGUCCGUCAAAAA 45% 14791 3UTR 2065 1803AUAGAAUGACAGUCCGUCAAAACAA 41% 14792 3UTR 2067 1804AGAAUGACAGUCCGUCAAAACAGAA 36% 14793 3UTR 2068 1805GAAUGACAGUCCGUCAAAACAGAUA 40% 14794 3UTR 2113 1806AGUGUCCUUGGCAGGCUGAUUUCUA 42% 14795 3UTR 2114 1807GUGUCCUUGGCAGGCUGAUUUCUAA 42% 14796 3UTR 2118 1808CCUUGGCAGGCUGAUUUCUAGGUAA 111% 14797 3UTR 2127 1809GCUGAUUUCUAGGUAGGAAAUGUGA 44% 14798 3UTR 2128 1810CUGAUUUCUAGGUAGGAAAUGUGGA 44% 14799 3UTR 2130 1811GAUUUCUAGGUAGGAAAUGUGGUAA 46% 14800 3UTR 2131 1812AUUUCUAGGUAGGAAAUGUGGUAGA 45% 14801 3UTR 2142 1813GGAAAUGUGGUAGCCUCACUUUUAA 37% 14802 3UTR 2146 1814AUGUGGUAGCCUCACUUUUAAUGAA 39% 14803 3UTR 2149 1815UGGUAGCCUCACUUUUAAUGAACAA 40% 14804 3UTR 2154 1816GCCUCACUUUUAAUGAACAAAUGGA 35% 14805 3UTR 2155 1817CCUCACUUUUAAUGAACAAAUGGCA 41% 14806 3UTR 2181 1818UUAUUAAAAACUGAGUGACUCUAUA 26% 14807 3UTR 2182 1819UAUUAAAAACUGAGUGACUCUAUAA 29% 14808 3UTR 2183 1820AUUAAAAACUGAGUGACUCUAUAUA 28% 14809 3UTR 2186 1821AAAAACUGAGUGACUCUAUAUAGCA 31% 14810 3UTR 2187 1822AAAACUGAGUGACUCUAUAUAGCUA 28% 14811 3UTR 2188 1823AAACUGAGUGACUCUAUAUAGCUGA 38% 14812 3UTR 2189 1824AACUGAGUGACUCUAUAUAGCUGAA 44% 14813 3UTR 2190 1825ACUGAGUGACUCUAUAUAGCUGAUA 38% 14814 3UTR 2255 1826ACUGUUUUUCGGACAGUUUAUUUGA 29% 14815 3UTR 2256 1827CUGUUUUUCGGACAGUUUAUUUGUA 25% 14816 3UTR 2263 1828UCGGACAGUUUAUUUGUUGAGAGUA 29% 14817 3UTR 2265 1829GGACAGUUUAUUUGUUGAGAGUGUA 24% 14818 3UTR 2268 1830CAGUUUAUUUGUUGAGAGUGUGACA 26% 14819 3UTR 2269 1831AGUUUAUUUGUUGAGAGUGUGACCA 37% 14820 3UTR 2272 1832UUAUUUGUUGAGAGUGUGACCAAAA 27% 14821 3UTR 2273 1833UAUUUGUUGAGAGUGUGACCAAAAA 30% 14822 3UTR 2274 1834AUUUGUUGAGAGUGUGACCAAAAGA 26% 14823 3UTR 2275 1835UUUGUUGAGAGUGUGACCAAAAGUA 27% 14824 3UTR 2276 1836UUGUUGAGAGUGUGACCAAAAGUUA 30% 14825 3UTR 2277 1837UGUUGAGAGUGUGACCAAAAGUUAA 29% 14826 3UTR 2278 1838GUUGAGAGUGUGACCAAAAGUUACA 33% 14827 3UTR 2279 1839UUGAGAGUGUGACCAAAAGUUACAA 35% 14828 3UTR 2281 1840GAGAGUGUGACCAAAAGUUACAUGA 36% 14829 3UTR 2282 1841AGAGUGUGACCAAAAGUUACAUGUA 36% 14830 3UTR 2283 1842GAGUGUGACCAAAAGUUACAUGUUA 33% 14831 3UTR 2284 1843AGUGUGACCAAAAGUUACAUGUUUA 31% 14832 3UTR 2285 1844GUGUGACCAAAAGUUACAUGUUUGA 22% 14833 3UTR 2286 1845UGUGACCAAAAGUUACAUGUUUGCA 40% 14834 3UTR 2293 1846AAAAGUUACAUGUUUGCACCUUUCA 24% 14835 3UTR 2295 1847AAGUUACAUGUUUGCACCUUUCUAA 23% 14836 3UTR 2296 1848AGUUACAUGUUUGCACCUUUCUAGA 29% 14837 3UTR 2299 1849UACAUGUUUGCACCUUUCUAGUUGA 27% 14838 3UTR 2300 1850ACAUGUUUGCACCUUUCUAGUUGAA 29% 14839 3UTR 2301 1851CAUGUUUGCACCUUUCUAGUUGAAA 35%

Key: PS * = Phosphothioate Backbone Linkage RNA G = Guanine RNA U =Uracil RNA C = Cytosine RNA A = Adenine m 2′ Omethyl f 2′-FluoroPhosphate P 5′ Phosphate

TABLE 12 Inhibition of gene expression with CTGF ori sequences(Accession Number: NM_001901.2) 25-mer Sense Strand (position 25 of SS,replaced with A) 25-mer Sense Strand (position 25 Oligo Gene Ref SEQ IDof SS, original base, A549 0.1 nM ID Region Pos NO not replaced by A)Activity 13843 CDS 1047 1852 UACCGAGCUAAAUUCUGUGGAGUAU 113% 13844 3UTR2164 1853 UAAUGAACAAAUGGCCUUUAUUAAA 61% 13845 3UTR 1795 1854UGGACAGCUUGUGGCAAGUGAAUUU 99% 13846 CDS 1228 1855UGUACUACAGGAAGAUGUACGGAGA 87% 13847 CDS 1146 1856GUCAUGAAGAAGAACAUGAUGUUCA 98% 13848 CDS 1150 1857UGAAGAAGAACAUGAUGUUCAUCAA 105% 13849 CDS 1218 1858UUUGAAUCGCUGUACUACAGGAAGA 91% 13850 3UTR 2262 1859UUCGGACAGUUUAUUUGUUGAGAGU 50% 13851 CDS 1147 1860UCAUGAAGAAGAACAUGAUGUUCAU 104% 13852 3UTR 2163 1861UUAAUGAACAAAUGGCCUUUAUUAA 54% 13853 3UTR 1414 1862CCAUGUCAAACAAAUAGUCUAUCAA 35% 13854 CDS 1195 1863ACUGUCCCGGAGACAAUGACAUCUU 103% 13855 3UTR 1788 1864AUUAGACUGGACAGCUUGUGGCAAG 103% 13856 3UTR 1793 1865ACUGGACAGCUUGUGGCAAGUGAAU 81% 13857 3UTR 1891 1866UAUAUAUGUACAGUUAUCUAAGUUA 73% 13858 3UTR 2270 1867GUUUAUUUGUUGAGAGUGUGACCAA 76% 13859 482 1868 CAAGAUCGGCGUGUGCACCGCCAAA95% 13860 CDS 942 1869 GCUGACCUGGAAGAGAACAUUAAGA 93% 13861 CDS 1199 1870UCCCGGAGACAAUGACAUCUUUGAA 83% 13862 3UTR 2258 1871GUUUUUCGGACAGUUUAUUUGUUGA 40% 13863 CDS 1201 1872CCGGAGACAAUGACAUCUUUGAAUC 123% 13864 CDS 543 1873CGCAGCGGAGAGUCCUUCCAGAGCA 124% 13865 3UTR 1496 1874AGCACCAGAAUGUAUAUUAAGGUGU 109% 13866 CDS 793 1875UGAUUAGAGCCAACUGCCUGGUCCA 125% 13867 CDS 1198 1876GUCCCGGAGACAAUGACAUCUUUGA 64% 13868 3UTR 2160 1877CUUUUAAUGAACAAAUGGCCUUUAU 68% 13869 CDS 1149 1878AUGAAGAAGAACAUGAUGUUCAUCA 107% 13870 CDS 1244 1879GUACGGAGACAUGGCAUGAAGCCAG 107% 13871 3UTR 1495 1880CAGCACCAGAAUGUAUAUUAAGGUG 77% 13872 475 1881 CCAACCGCAAGAUCGGCGUGUGCAC113% 13873 CDS 806 1882 CUGCCUGGUCCAGACCACAGAGUGG 113% 13874 CDS 8191883 ACCACAGAGUGGAGCGCCUGUUCCA 99% 13875 CDS 1221 1884GAAUCGCUGUACUACAGGAAGAUGU 97% 13876 CDS 1152 1885AAGAAGAACAUGAUGUUCAUCAAGA 121% 13877 CDS 1163 1886GAUGUUCAUCAAGACCUGUGCCUGC 125% 13878 3UTR 1494 1887ACAGCACCAGAAUGUAUAUUAAGGU 94% 13879 3UTR 1890 1888AUAUAUAUGUACAGUUAUCUAAGUU 94% 13880 473 1889 GGCCAACCGCAAGAUCGGCGUGUGC122% 13881 544 1890 GCAGCGGAGAGUCCUUCCAGAGCAG 111% 13882 CDS 883 1891ACAACGCCUCCUGCAGGCUAGAGAA 105% 13883 CDS 1240 1892AGAUGUACGGAGACAUGGCAUGAAG 99% 13884 CDS 1243 1893UGUACGGAGACAUGGCAUGAAGCCA 116% 13885 3UTR 2266 1894GACAGUUUAUUUGUUGAGAGUGUGA 53% 13886 CDS 1011 1895AAGUUUGAGCUUUCUGGCUGCACCA 118% 13887 CDS 1020 1896CUUUCUGGCUGCACCAGCAUGAAGA 110% 13888 CDS 1168 1897UCAUCAAGACCUGUGCCUGCCAUUA 119% 13889 1415 1898 CAUGUCAAACAAAUAGUCUAUCAAC64% 13890 3UTR 1792 1899 GACUGGACAGCUUGUGGCAAGUGAA 53% 13891 3UTR 21561900 CUCACUUUUAAUGAACAAAUGGCCU 119% 13892 379 1901GCUGCCGCGUCUGCGCCAAGCAGCU 112% 13893 CDS 1229 1902GUACUACAGGAAGAUGUACGGAGAC 112% 13894 3UTR 1791 1903AGACUGGACAGCUUGUGGCAAGUGA 65% 13895 3UTR 2158 1904CACUUUUAAUGAACAAAUGGCCUUU 76% 13896 488 1905 CGGCGUGUGCACCGCCAAAGAUGGU89% 13897 CDS 1151 1906 GAAGAAGAACAUGAUGUUCAUCAAG 119% 13898 CDS 11561907 AGAACAUGAUGUUCAUCAAGACCUG 125% 13899 CDS 1237 1908GGAAGAUGUACGGAGACAUGGCAUG 114% 13900 CDS 1202 1909CGGAGACAAUGACAUCUUUGAAUCG 130% 13901 CDS 1236 1910AGGAAGAUGUACGGAGACAUGGCAU 135% 13902 3UTR 1786 1911CGAUUAGACUGGACAGCUUGUGGCA 119% 13903 3UTR 1789 1912UUAGACUGGACAGCUUGUGGCAAGU 108% 13904 3UTR 2290 1913ACCAAAAGUUACAUGUUUGCACCUU 90% 13905 CDS 1017 1914GAGCUUUCUGGCUGCACCAGCAUGA 121% 13906 CDS 1197 1915UGUCCCGGAGACAAUGACAUCUUUG 125% 13907 CDS 1219 1916UUGAAUCGCUGUACUACAGGAAGAU 98% 13908 3UTR 2159 1917ACUUUUAAUGAACAAAUGGCCUUUA 52% 13909 486 1918 AUCGGCGUGUGCACCGCCAAAGAUG119% 13910 CDS 826 1919 AGUGGAGCGCCUGUUCCAAGACCUG 139% 13911 CDS 10221920 UUCUGGCUGCACCAGCAUGAAGACA 144% 13912 3UTR 1492 1921ACACAGCACCAGAAUGUAUAUUAAG 99% 13913 3UTR 1781 1922CCUGUCGAUUAGACUGGACAGCUUG 89% 13914 485 1923 GAUCGGCGUGUGCACCGCCAAAGAU131% 13915 CDS 1007 1924 UAUCAAGUUUGAGCUUUCUGGCUGC 92% 13916 CDS 12421925 AUGUACGGAGACAUGGCAUGAAGCC 106% 13917 3UTR 1787 1926GAUUAGACUGGACAGCUUGUGGCAA 104% 13918 3UTR 1889 1927UAUAUAUAUGUACAGUUAUCUAAGU 78% 13919 3UTR 2294 1928AAAGUUACAUGUUUGCACCUUUCUA 28% 13920 CDS 821 1929CACAGAGUGGAGCGCCUGUUCCAAG 108% 13921 CDS 884 1930CAACGCCUCCUGCAGGCUAGAGAAG 125% 13922 3UTR 2260 1931UUUUCGGACAGUUUAUUUGUUGAGA 43% 13923 CDS 889 1932CCUCCUGCAGGCUAGAGAAGCAGAG 95% 13924 CDS 1226 1933GCUGUACUACAGGAAGAUGUACGGA 122% 13925 3UTR 1493 1934CACAGCACCAGAAUGUAUAUUAAGG 88% 13926 3UTR 1799 1935CAGCUUGUGGCAAGUGAAUUUGCCU 89% 13927 CDS 807 1936UGCCUGGUCCAGACCACAGAGUGGA 101% 13928 CDS 1107 1937ACCACCCUGCCGGUGGAGUUCAAGU 113% 13929 CDS 1155 1938AAGAACAUGAUGUUCAUCAAGACCU 109% 13930 CDS 1169 1939CAUCAAGACCUGUGCCUGCCAUUAC 89% 13931 CDS 1241 1940GAUGUACGGAGACAUGGCAUGAAGC 96% 13932 3UTR 1794 1941CUGGACAGCUUGUGGCAAGUGAAUU 73% 13933 3UTR 1888 1942AUAUAUAUAUGUACAGUUAUCUAAG 98% 13934 3UTR 2289 1943GACCAAAAGUUACAUGUUUGCACCU 77% 13935 373 1944 GCGGCUGCUGCCGCGUCUGCGCCAA85% 13936 CDS 799 1945 GAGCCAACUGCCUGGUCCAGACCAC 126% 13937 CDS 802 1946CCAACUGCCUGGUCCAGACCACAGA 122% 13938 CDS 1166 1947GUUCAUCAAGACCUGUGCCUGCCAU 106%

TABLE 13 Inhibition of gene expression with SPP1 sd-rxRNA sequences(Accession Number: NM_000582.2) % remaining Oligo Start SEQ ID SEQ IDexpression (1 Number Site NO Sense sequence NO Antisense sequence uMA549) 14084 1025 1948 CUCAUGAAUUAGA 1949 UCUAAUUCAUGAGAA 61% AUAC 140851049 1950 CUGAGGUCAAUUA 1951 UAAUUGACCUCAGAA 50% GAUG 14086 1051 1952GAGGUCAAUUAAA 1953 UUUAAUUGACCUCAG n/a AAGA 14087 1048 1954UCUGAGGUCAAUU 1955 AAUUGACCUCAGAAG 69% AUGC 14088 1050 1956UGAGGUCAAUUAA 1957 UUAAUUGACCUCAGA 76% AGAU 14089 1047 1958UUCUGAGGUCAAU 1959 AUUGACCUCAGAAGA 60% UGCA 14090 800 1960 GUCAGCUGGAUGA1961 UCAUCCAGCUGACUC 71% GUUU 14091 492 1962 UUCUGAUGAAUCU 1963AGAUUCAUCAGAAUG n/a GUGA 14092 612 1964 UGGACUGAGGUCA 1965UGACCUCAGUCCAUA n/a AACC 14093 481 1966 GAGUCUCACCAUU 1967AAUGGUGAGACUCAU n/a CAGA 14094 614 1968 GACUGAGGUCAAA 1969UUUGACCUCAGUCCA n/a UAAA 14095 951 1970 UCACAGCCAUGAA 1971UUCAUGGCUGUGAAA 89% UUCA 14096 482 1972 AGUCUCACCAUUC 1973GAAUGGUGAGACUCA 87% UCAG 14097 856 1974 AAGCGGAAAGCCA 1975UGGCUUUCCGCUUAU 88% AUAA 14098 857 1976 AGCGGAAAGCCAA 1977UUGGCUUUCCGCUUA 113% UAUA 14099 365 1978 ACCACAUGGAUGA 1979UCAUCCAUGUGGUCA 98% UGGC 14100 359 1980 GCCAUGACCACAU 1981AUGUGGUCAUGGCU 84% UUCGU 14101 357 1982 AAGCCAUGACCAC 1983GUGGUCAUGGCUUU 88% CGUUG 14102 858 1984 GCGGAAAGCCAAU 1985AUUGGCUUUCCGCUU n/a AUAU 14103 1012 1986 AAAUUUCGUAUUU 1987AAAUACGAAAUUUCA 93% GGUG 14104 1014 1988 AUUUCGUAUUUCU 1989AGAAAUACGAAAUUU 89% CAGG 14105 356 1990 AAAGCCAUGACCA 1991UGGUCAUGGCUUUC 85% GUUGG 14106 368 1992 ACAUGGAUGAUAU 1993AUAUCAUCCAUGUGG 67% UCAU 14107 1011 1994 GAAAUUUCGUAUU 1995AAUACGAAAUUUCAG 87% GUGU 14108 754 1996 GCGCCUUCUGAUU 1997AAUCAGAAGGCGCGU 73% UCAG 14109 1021 1998 AUUUCUCAUGAAU 1999AUUCAUGAGAAAUAC 128% GAAA 14110 1330 2000 CUCUCAUGAAUAG 2001CUAUUCAUGAGAGAA 101% UAAC 14111 346 2002 AAGUCCAACGAAA 2003UUUCGUUGGACUUA 59% CUUGG 14112 869 2004 AUGAUGAGAGCAA 2005UUGCUCUCAUCAUUG 89% GCUU 14113 701 2006 GCGAGGAGUUGAA 2007UUCAACUCCUCGCUU 95% UCCA 14114 896 2008 UGAUUGAUAGUCA 2009UGACUAUCAAUCACA 87% UCGG 14115 1035 2010 AGAUAGUGCAUCU 2011AGAUGCACUAUCUAA 82% UUCA 14116 1170 2012 AUGUGUAUCUAUU 2013AAUAGAUACACAUUC 36% AACC 14117 1282 2014 UUCUAUAGAAGAA 2015UUCUUCUAUAGAAU 91% GAACA 14118 1537 2016 UUGUCCAGCAAUU 2017AAUUGCUGGACAACC 152% GUGG 14119 692 2018 ACAUGGAAAGCGA 2019UCGCUUUCCAUGUGU n/a GAGG 14120 840 2020 GCAGUCCAGAUUA 2021UAAUCUGGACUGCUU 87% GUGG 14121 1163 2022 UGGUUGAAUGUGU 2023ACACAUUCAACCAAU 31% AAAC 14122 789 2024 UUAUGAAACGAGU 2025ACUCGUUUCAUAACU 96% GUCC 14123 841 2026 CAGUCCAGAUUAU 2027AUAAUCUGGACUGCU 110% UGUG 14124 852 2028 AUAUAAGCGGAAA 2029UUUCCGCUUAUAUAA 91% UCUG 14125 209 2030 UACCAGUUAAACA 2031UGUUUAACUGGUAU 110% GGCAC 14126 1276 2032 UGUUCAUUCUAUA 2033UAUAGAAUGAACAUA n/a GACA 14127 137 2034 CCGACCAAGGAAA 2035UUUCCUUGGUCGGC 71% GUUUG 14128 711 2036 GAAUGGUGCAUAC 2037GUAUGCACCAUUCAA 115% CUCC 14129 582 2038 AUAUGAUGGCCGA 2039UCGGCCAUCAUAUGU 97% GUCU 14130 839 2040 AGCAGUCCAGAUU 2041AAUCUGGACUGCUUG 102% UGGC 14131 1091 2042 GCAUUUAGUCAAA 2043UUUGACUAAAUGCAA 10% AGUG 14132 884 2044 AGCAUUCCGAUGU 2045ACAUCGGAAUGCUCA 93% UUGC 14133 903 2046 UAGUCAGGAACUU 2047AAGUUCCUGACUAUC 97% AAUC 14134 1090 2048 UGCAUUUAGUCAA 2049UUGACUAAAUGCAAA 39% GUGA 14135 474 2050 GUCUGAUGAGUCU 2051AGACUCAUCAGACUG 99% GUGA 14136 575 2052 UAGACACAUAUGA 2053UCAUAUGUGUCUACU 108% GUGG 14137 671 2054 CAGACGAGGACAU 2055AUGUCCUCGUCUGUA 98% GCAU 14138 924 2056 CAGCCGUGAAUUC 2057GAAUUCACGGCUGAC 100% UUUG 14139 1185 2058 AGUCUGGAAAUAA 2059UUAUUUCCAGACUCA 47% AAUA 14140 1221 2060 AGUUUGUGGCUUC 2061GAAGCCACAAACUAA 100% ACUA 14141 347 2062 AGUCCAACGAAAG 2063CUUUCGUUGGACUU 103% ACUUG 14142 634 2064 AAGUUUCGCAGAC 2065GUCUGCGAAACUUCU 100% UAGA 14143 877 2066 AGCAAUGAGCAUU 2067AAUGCUCAUUGCUCU 104% CAUC 14144 1033 2068 UUAGAUAGUGCAU 2069AUGCACUAUCUAAUU 95% CAUG 14145 714 2070 UGGUGCAUACAAG 2071CUUGUAUGCACCAUU 101% CAAC 14146 791 2072 AUGAAACGAGUCA 2073UGACUCGUUUCAUAA 100% CUGU 14147 813 2074 CCAGAGUGCUGAA 2075UUCAGCACUCUGGUC 97% AUCC 14148 939 2076 CAGCCAUGAAUUU 2077AAAUUCAUGGCUGUG 109% GAAU 14149 1161 2078 AUUGGUUGAAUGU 2079ACAUUCAACCAAUAA 34% ACUG 14150 1164 2080 GGUUGAAUGUGUA 2081UACACAUUCAACCAA n/a UAAA 14151 1190 2082 GGAAAUAACUAAU 2083AUUAGUUAUUUCCA n/a GACUC 14152 1333 2084 UCAUGAAUAGAAA 2085UUUCUAUUCAUGAG 31% AGAAU 14153 537 2086 GCCAGCAACCGAA 2087UUCGGUUGCUGGCA n/a GGUCC 14154 684 2088 CACCUCACACAUG 2089CAUGUGUGAGGUGA 100% UGUCC 14155 707 2090 AGUUGAAUGGUGC 2091GCACCAUUCAACUCC 99% UCGC 14156 799 2092 AGUCAGCUGGAUG 2093CAUCCAGCUGACUCG 95% UUUC 14157 853 2094 UAUAAGCGGAAAG 2095CUUUCCGCUUAUAUA 106% AUCU 14158 888 2096 UUCCGAUGUGAUU 2097AAUCACAUCGGAAUG 88% CUCA 14159 1194 2098 AUAACUAAUGUGU 2099ACACAUUAGUUAUUU 95% CCAG 14160 1279 2100 UCAUUCUAUAGAA 2101UUCUAUAGAAUGAAC 15% AUAG 14161 1300 2102 AACUAUCACUGUA 2103UACAGUGAUAGUUU 86% GCAUU 14162 1510 2104 GUCAAUUGCUUAU 2105AUAAGCAAUUGACAC 86% CACC 14163 1543 2106 AGCAAUUAAUAAA 2107UUUAUUAAUUGCUG 110% GACAA 14164 434 2108 ACGACUCUGAUGA 2109UCAUCAGAGUCGUUC 134% GAGU 14165 600 2110 UAGUGUGGUUUAU 2111AUAAACCACACUAUC 102% ACCU 14166 863 2112 AAGCCAAUGAUGA 2113UCAUCAUUGGCUUUC 93% CGCU 14167 902 2114 AUAGUCAGGAACU 2115AGUUCCUGACUAUCA 101% AUCA 14168 921 2116 AGUCAGCCGUGAA 2117UUCACGGCUGACUUU 98% GGAA 14169 154 2118 ACUACCAUGAGAA 2119UUCUCAUGGUAGUG n/a AGUUU 14170 217 2120 AAACAGGCUGAUU 2121AAUCAGCCUGUUUAA 66% CUGG 14171 816 2122 GAGUGCUGAAACC 2123GGUUUCAGCACUCUG 102% GUCA 14172 882 2124 UGAGCAUUCCGAU 2125AUCGGAAUGCUCAUU 103% GCUC 14173 932 2126 AAUUCCACAGCCA 2127UGGCUGUGGAAUUC n/a ACGGC 14174 1509 2128 UGUCAAUUGCUUA 2129UAAGCAAUUGACACC n/a ACCA 14175 157 2130 ACCAUGAGAAUUG 2131CAAUUCUCAUGGUAG 109% UGAG 14176 350 2132 CCAACGAAAGCCA 2133UGGCUUUCGUUGGA 95% CUUAC 14177 511 2134 CUGGUCACUGAUU 2135AAUCAGUGACCAGUU 100% CAUC 14178 605 2136 UGGUUUAUGGACU 2137AGUCCAUAAACCACA 99% CUAU 14179 811 2138 GACCAGAGUGCUG 2139CAGCACUCUGGUCAU 88% CCAG 14180 892 2140 GAUGUGAUUGAUA 2141UAUCAAUCACAUCGG 76% AAUG 14181 922 2142 GUCAGCCGUGAAU 2143AUUCACGGCUGACUU 59% UGGA 14182 1169 2144 AAUGUGUAUCUAU 2145AUAGAUACACAUUCA 69% ACCA 14183 1182 2146 UUGAGUCUGGAAA 2147UUUCCAGACUCAAAU n/a AGAU 14184 1539 2148 GUCCAGCAAUUAA 2149UUAAUUGCUGGACAA 77% CCGU 14185 1541 2150 CCAGCAAUUAAUA 2151UAUUAAUUGCUGGA n/a CAACC 14186 427 2152 GACUCGAACGACU 2153AGUCGUUCGAGUCAA 69% UGGA 14187 533 2154 ACCUGCCAGCAAC 2155GUUGCUGGCAGGUCC 78% GUGG 18538 496 2156 GAUGAAUCUGAUA 2157UAUCAGAUUCAUCAG 74% AAUG 18539 496 2158 UGAUGAAUCUGA 2159UAUCAGAUUCAUCAG 72% UA AAUG 18540 175 2160 AUUUGCUUUUGCA 2161UGCAAAAGCAAAUCA 98% CUGC 18541 175 2162 GAUUUGCUUUUG 2163UGCAAAAGCAAAUCA 28% CA CUGC 18542 172 2164 GUGAUUUGCUUUA 2165UAAAGCAAAUCACUG 24% CAAU 18543 172 2166 AGUGAUUUGCUU 2167UAAAGCAAAUCACUG 14% UA CAAU 18544 1013 2168 AAUUUCGUAUUUA 2169UAAAUACGAAAUUUC 100% AGGU 18545 1013 2170 AAAUUUCGUAUU 2171UAAAUACGAAAUUUC 109% UA AGGU 18546 952 2172 CACAGCCAUGAAA 2173UUUCAUGGCUGUGA 32% AAUUC 18547 952 2174 UCACAGCCAUGAAA 2175UUUCAUGGCUGUGA 33% AAUUC 18548 174 2176 GAUUUGCUUUUGA 2177UCAAAAGCAAAUCAC 57% UGCA 18549 174 2178 UGAUUUGCUUUU 2179UCAAAAGCAAAUCAC 53% GA UGCA 18550 177 2180 UUGCUUUUGCCUA 2181UAGGCAAAAGCAAAU 97% CACU 18551 177 2182 UUUGCUUUUGCC 2183UAGGCAAAAGCAAAU 103% UA CACU 18552 1150 2184 UUUCUCAGUUUAA 2185UUAAACUGAGAAAGA 96% AGCA 18553 1089 2186 UUGCAUUUAGUCA 2187UGACUAAAUGCAAAG 94% UGAG 18554 1086 2188 ACUUUGCAUUUAA 2189UUAAAUGCAAAGUGA n/a GAAA 18555 1093 2190 AUUUAGUCAAAAA 2191UUUUUGACUAAAUG n/a CAAAG 18556 1147 2192 UUCUUUCUCAGUA 2193UACUGAGAAAGAAGC n/a AUUU 18557 1148 2194 UCUUUCUCAGUUA 2195UAACUGAGAAAGAAG 66% CAUU 18558 1128 2196 GAAAGAGAACAUA 2197UAUGUUCUCUUUCA 16% UUUUG 18559 1087 2198 CUUUGCAUUUAGA 2199UCUAAAUGCAAAGUG 28% AGAA 18560 1088 2200 UUUGCAUUUAGUA 2201UACUAAAUGCAAAGU n/a GAGA 18561 1083 2202 CUCACUUUGCAUA 2203UAUGCAAAGUGAGAA 53% AUUG 18562 1081 2204 UUCUCACUUUGCA 2205UGCAAAGUGAGAAAU 89% UGUA 18563 555 2206 CACUCCAGUUGUA 2207UACAACUGGAGUGAA 33% AACU 18564 1125 2208 AAUGAAAGAGAAA 2209UUUCUCUUUCAUUU n/a UGCUA 18565 168 2210 UGCAGUGAUUUGA 2211UCAAAUCACUGCAAU 14% UCUC 18566 1127 2212 UGAAAGAGAACAA 2213UUGUUCUCUUUCAU 27% UUUGC 18567 1007 2214 ACCUGAAAUUUCA 2215UGAAAUUUCAGGUG 129% UUUAU 18568 164 2216 GAAUUGCAGUGAA 2217UUCACUGCAAUUCUC 47% AUGG 18569 222 2218 GGCUGAUUCUGGA 2219UCCAGAAUCAGCCUG n/a UUUA

TABLE 14 Inhibition of gene expression with PTGS2 sd-rxRNA sequences(Accession Number: NM_000963.2) Oligo Start SEQ ID SEQ ID Number Site NOSense sequence NO Antisense sequence % remaining expression (1 uM A549)14422 451 2220 CACAUUUGAUUGA 2221 UCAAUCAAAUGUGAUC 72% UGG 14423 17692222 CACUGCCUCAAUU 2223 AAUUGAGGCAGUGUU 71% GAUG 14424 1464 2224AAAUACCAGUCUU 2225 AAGACUGGUAUUUCAU 74% CUG 14425 453 2226 CAUUUGAUUGACA2227 UGUCAAUCAAAUGUGA 83% UCU % remaining expression (1 uM PC-3) 17388285 2228 GAAAACUGCUCAA 2229 UUGAGCAGUUUUCUCC 88% AUA 17389 520 2230ACCUCUCCUAUUA 2231 UAAUAGGAGAGGUUA 25% GAGA 17390 467 2232 UCCACCAACUUAA2233 UUAAGUUGGUGGACU 68% GUCA 17391 467 2234 GUCCACCAACUUAA 2235UUAAGUUGGUGGACU 101% GUCA 17392 524 2236 CUCCUAUUAUACA 2237UGUAUAAUAGGAGAG 49% GUUA 17393 448 2238 GAUCACAUUUGAA 2239UUCAAAUGUGAUCUG 29% GAUG 17394 448 2240 AGAUCACAUUUGAA 2241UUCAAAUGUGAUCUG 31% GAUG 17395 519 2242 AACCUCUCCUAUA 2243UAUAGGAGAGGUUAG 12% AGAA 17396 437 2244 GUUGACAUCCAGA 2245UCUGGAUGUCAACACA 86% UAA 17397 406 2246 CCUUCCUUCGAAA 2247UUUCGAAGGAAGGGAA 23% UGU 17398 339 2248 ACUCCAAACACAA 2249UUGUGUUUGGAGUGG 102% GUUU 17399 339 2250 CACUCCAAACACAA 2251UUGUGUUUGGAGUGG 55% GUUU 17400 338 2252 CACUCCAAACACA 2253UGUGUUUGGAGUGGG 62% UUUC 17401 468 2254 CCACCAACUUACA 2255UGUAAGUUGGUGGAC 61% UGUC 17402 468 2256 UCCACCAACUUACA 2257UGUAAGUUGGUGGAC 179% UGUC 17403 1465 2258 AAUACCAGUCUUA 2259UAAGACUGGUAUUUCA 30% UCU 17404 243 2260 GACCAGUAUAAGA 2261UCUUAUACUGGUCAAA 32% UCC 17405 1472 2262 GUCUUUUAAUGAA 2263UUCAUUAAAAGACUGG 15% UAU 17406 2446 2264 AAUUUCAUGUCUA 2265UAGACAUGAAAUUACU 142% GGU 17407 449 2266 AUCACAUUUGAUA 2267UAUCAAAUGUGAUCUG 54% GAU 17408 449 2268 GAUCACAUUUGAUA 2269UAUCAAAUGUGAUCUG 27% GAU 17409 444 2270 UCCAGAUCACAUA 2271UAUGUGAUCUGGAUG 49% UCAA 17410 1093 2272 UACUGAUAGGAGA 2273UCUCCUAUCAGUAUUA 32% GCC 17411 1134 2274 GUGCAACACUUGA 2275UCAAGUGUUGCACAUA 70% AUC 17412 244 2276 ACCAGUAUAAGUA 2277UACUUAUACUGGUCAA 63% AUC 17413 1946 2278 GAAGUCUAAUGAA 2279UUCAUUAGACUUCUAC 19% AGU 17414 638 2280 AAGAAGAAAGUUA 2281UAACUUUCUUCUUAGA 27% AGC 17415 450 2282 UCACAUUUGAUUA 2283UAAUCAAAUGUGAUCU 216% GGA 17416 450 2284 AUCACAUUUGAUUA 2285UAAUCAAAUGUGAUCU 32% GGA 17417 452 2286 ACAUUUGAUUGAA 2287UUCAAUCAAAUGUGAU 99% CUG 17418 452 2288 CACAUUUGAUUGAA 2289UUCAAUCAAAUGUGAU 54% CUG 17419 454 2290 AUUUGAUUGACAA 2291UUGUCAAUCAAAUGUG 86% AUC 17420 454 2292 CAUUUGAUUGACAA 2293UUGUCAAUCAAAUGUG 89% AUC 17421 1790 2294 CAUCUGCAAUAAA 2295UUUAUUGCAGAUGAG 55% AGAC 17422 1790 2296 UCAUCUGCAAUAAA 2297UUUAUUGCAGAUGAG 62% AGAC

TABLE 15 Inhibition of gene expression with CTGF sd-rxRNA sequences(Accession number: NM_001901.2) % remaining mRNA Oligo Start SEQ ID SEQID expression (1 uM Number Site NO Sense sequence NO Antisense sequencesd-rxRNA, A549) 13980 1222 2298 ACAGGAAGAUG 2299 UACAUCUUCCUGUAG 98% UAUACA 13981 813 2300 GAGUGGAGCGC 2301 AGGCGCUCCACUCUG 82% CU UGGU 13982747 2302 CGACUGGAAGA 4206 UGUCUUCCAGUCGGU 116% CA AAGC 13983 817 2303GGAGCGCCUGU 4207 GAACAGGCGCUCCAC 97% UC UCUG 13984 1174 2304 GCCAUUACAAC4208 CAGUUGUAAUGGCAG 102% UG GCAC 13985 1005 2305 GAGCUUUCUG 4209AGCCAGAAAGCUCAA 114% GCU ACUU 13986 814 2306 AGUGGAGCGCC 4210CAGGCGCUCCACUCU 111% UG GUGG 13987 816 2307 UGGAGCGCCUG 4211AACAGGCGCUCCACU 102% UU CUGU 13988 1001 2308 GUUUGAGCUU 4212AGAAAGCUCAAACUU 99% UCU GAUA 13989 1173 2309 UGCCAUUACAA 4213AGUUGUAAUGGCAG 107% CU GCACA 13990 749 2310 ACUGGAAGACA 4214CGUGUCUUCCAGUCG 91% CG GUAA 13991 792 2311 AACUGCCUGGU 4215GGACCAGGCAGUUGG 97% CC CUCU 13992 1162 2312 AGACCUGUGCC 4216CAGGCACAGGUCUUG 107% UG AUGA 13993 811 2313 CAGAGUGGAGC 4217GCGCUCCACUCUGUG 113% GC GUCU 13994 797 2314 CCUGGUCCAGA 4218GGUCUGGACCAGGCA n/a CC GUUG 13995 1175 2315 CCAUUACAACU 4219ACAGUUGUAAUGGCA 113% GU GGCA 13996 1172 2316 CUGCCAUUACA 4220GUUGUAAUGGCAGG 110% AC CACAG 13997 1177 2317 AUUACAACUGU 4221GGACAGUUGUAAUG 105% CC GCAGG 13998 1176 2318 CAUUACAACUG 4222GACAGUUGUAAUGGC 89% UC AGGC 13999 812 2319 AGAGUGGAGCG 4223GGCGCUCCACUCUGU 99% CC GGUC 14000 745 2320 ACCGACUGGAA 4224UCUUCCAGUCGGUAA n/a GA GCCG 14001 1230 2321 AUGUACGGAGA 4225UGUCUCCGUACAUCU 106% CA UCCU 14002 920 2322 GCCUUGCGAAG 4226AGCUUCGCAAGGCCU 93% CU GACC 14003 679 2323 GCUGCGAGGAG 4227CACUCCUCGCAGCAU 102% UG UUCC 14004 992 2324 GCCUAUCAAGU 4228AAACUUGAUAGGCUU 100% UU GGAG 14005 1045 2325 AAUUCUGUGG 4229ACUCCACAGAAUUUA 104% AGU GCUC 14006 1231 2326 UGUACGGAGAC 4230AUGUCUCCGUACAUC 87% AU UUCC 14007 991 2327 AGCCUAUCAAG 4231AACUUGAUAGGCUUG 101% UU GAGA 14008 998 2328 CAAGUUUGAGC 4232AAGCUCAAACUUGAU 98% UU AGGC 14009 1049 2329 CUGUGGAGUA 4233ACAUACUCCACAGAA 98% UGU UUUA 14010 1044 2330 AAAUUCUGUG 4234CUCCACAGAAUUUAG 93% GAG CUCG 14011 1327 2331 UUUCAGUAGCA 4235UGUGCUACUGAAAUC 95% CA AUUU 14012 1196 2332 CAAUGACAUCU 4236AAAGAUGUCAUUGUC 101% UU UCCG 14013 562 2333 AGUACCAGUGC 4237GUGCACUGGUACUUG 66% AC CAGC 14014 752 2334 GGAAGACACGU 4238AAACGUGUCUUCCAG 95% UU UCGG 14015 994 2335 CUAUCAAGUUU 4239UCAAACUUGAUAGGC 85% GA UUGG 14016 1040 2336 AGCUAAAUUCU 4240ACAGAAUUUAGCUCG 61% GU GUAU 14017 1984 2337 AGGUAGAAUG 4241UUACAUUCUACCUAU 32% UAA GGUG 14018 2195 2338 AGCUGAUCAGU 4242AAACUGAUCAGCUAU 86% UU AUAG 14019 2043 2339 UUCUGCUCAGA 4243UAUCUGAGCAGAAUU 81% UA UCCA 14020 1892 2340 UUAUCUAAGU 4244UUAACUUAGAUAACU 84% UAA GUAC 14021 1567 2341 UAUACGAGUAA 4245UAUUACUCGUAUAAG 72% UA AUGC 14022 1780 2342 GACUGGACAGC 4246AAGCUGUCCAGUCUA 65% UU AUCG 14023 2162 2343 AUGGCCUUUAU 4247UAAUAAAGGCCAUUU 80% UA GUUC 14024 1034 2344 AUACCGAGCUA 4248UUUAGCUCGGUAUG 91% AA UCUUC 14025 2264 2345 UUGUUGAGAG 4249ACACUCUCAACAAAU 58% UGU AAAC 14026 1032 2346 ACAUACCGAGC 4250UAGCUCGGUAUGUC 106% UA UUCAU 14027 1535 2347 AGCAGAAAGGU 4251UAACCUUUCUGCUGG 67% UA UACC 14028 1694 2348 AGUUGUUCCU 4252UUAAGGAACAACUUG 94% UAA ACUC 14029 1588 2349 AUUUGAAGUG 4253UUACACUUCAAAUAG 97% UAA CAGG 14030 928 2350 AAGCUGACCUG 4254UCCAGGUCAGCUUCG 100% GA CAAG 14031 1133 2351 GGUCAUGAAGA 4255CUUCUUCAUGACCUC 82% AG GCCG 14032 912 2352 AUGGUCAGGCC 4256AAGGCCUGACCAUGC 84% UU ACAG 14033 753 2353 GAAGACACGUU 4257CAAACGUGUCUUCCA 86% UG GUCG 14034 918 2354 AGGCCUUGCGA 4258CUUCGCAAGGCCUGA 88% AG CCAU 14035 744 2355 UACCGACUGGA 4259CUUCCAGUCGGUAAG 95% AG CCGC 14036 466 2356 ACCGCAAGAUC 4260CCGAUCUUGCGGUUG 73% GG GCCG 14037 917 2357 CAGGCCUUGCG 4261UUCGCAAGGCCUGAC 86% AA CAUG 14038 1038 2358 CGAGCUAAAUU 4262AGAAUUUAGCUCGGU 84% CU AUGU 14039 1048 2359 UCUGUGGAGU 4263CAUACUCCACAGAAU 87% AUG UUAG 14040 1235 2360 CGGAGACAUGG 4264UGCCAUGUCUCCGUA 100% CA CAUC 14041 868 2361 AUGACAACGCC 4265GAGGCGUUGUCAUU 104% UC GGUAA 14042 1131 2362 GAGGUCAUGAA 4266UCUUCAUGACCUCGC 85% GA CGUC 14043 1043 2363 UAAAUUCUGU 4267UCCACAGAAUUUAGC 74% GGA UCGG 14044 751 2364 UGGAAGACACG 4268AACGUGUCUUCCAGU 84% UU CGGU 14045 1227 2365 AAGAUGUACGG 4269CUCCGUACAUCUUCC 99% AG UGUA 14046 867 2366 AAUGACAACGC 4270AGGCGUUGUCAUUG 94% CU GUAAC 14047 1128 2367 GGCGAGGUCAU 4271UCAUGACCUCGCCGU 89% GA CAGG 14048 756 2368 GACACGUUUGG 4272GGCCAAACGUGUCUU 93% CC CCAG 14049 1234 2369 ACGGAGACAUG 4273GCCAUGUCUCCGUAC 100% GC AUCU 14050 916 2370 UCAGGCCUUGC 4274UCGCAAGGCCUGACC 96% GA AUGC 14051 925 2371 GCGAAGCUGAC 4275AGGUCAGCUUCGCAA 80% CU GGCC 14052 1225 2372 GGAAGAUGUAC 4276CCGUACAUCUUCCUG 96% GG UAGU 14053 445 2373 GUGACUUCGGC 4277GAGCCGAAGUCACAG 101% UC AAGA 14054 446 2374 UGACUUCGGCU 4278GGAGCCGAAGUCACA 93% CC GAAG 14055 913 2375 UGGUCAGGCCU 4279CAAGGCCUGACCAUG 67% UG CACA 14056 997 2376 UCAAGUUUGA 4280AGCUCAAACUUGAUA 92% GCU GGCU 14057 277 2377 GCCAGAACUGC 4281CUGCAGUUCUGGCCG 84% AG ACGG 14058 1052 2378 UGGAGUAUGU 4282GGUACAUACUCCACA n/a ACC GAAU 14059 887 2379 GCUAGAGAAGC 4283CUGCUUCUCUAGCCU 80% AG GCAG 14060 914 2380 GGUCAGGCCUU 4284GCAAGGCCUGACCAU 112% GC GCAC 14061 1039 2381 GAGCUAAAUUC 4285CAGAAUUUAGCUCGG 104% UG UAUG 14062 754 2382 AAGACACGUUU 4286CCAAACGUGUCUUCC 109% GG AGUC 14063 1130 2383 CGAGGUCAUGA 4287CUUCAUGACCUCGCC 103% AG GUCA 14064 919 2384 GGCCUUGCGAA 4288GCUUCGCAAGGCCUG 109% GC ACCA 14065 922 2385 CUUGCGAAGCU 4289UCAGCUUCGCAAGGC 106% GA CUGA 14066 746 2386 CCGACUGGAAG 4290GUCUUCCAGUCGGUA 106% AC AGCC 14067 993 2387 CCUAUCAAGUU 4291CAAACUUGAUAGGCU 67% UG UGGA 14068 825 2388 UGUUCCAAGAC 4292AGGUCUUGGAACAGG 93% CU CGCU 14069 926 2389 CGAAGCUGACC 4293CAGGUCAGCUUCGCA 95% UG AGGC 14070 923 2390 UUGCGAAGCUG 4294GUCAGCUUCGCAAGG 95% AC CCUG 14071 866 2391 CAAUGACAACG 4295GGCGUUGUCAUUGG 132% CC UAACC 14072 563 2392 GUACCAGUGCA 4296CGUGCACUGGUACUU n/a CG GCAG 14073 823 2393 CCUGUUCCAAG 4297GUCUUGGAACAGGCG 98% AC CUCC 14074 1233 2394 UACGGAGACAU 4298CCAUGUCUCCGUACA 109% GG UCUU 14075 924 2395 UGCGAAGCUGA 4299GGUCAGCUUCGCAAG 95% CC GCCU 14076 921 2396 CCUUGCGAAGC 4300CAGCUUCGCAAGGCC 116% UG UGAC 14077 443 2397 CUGUGACUUCG 4301GCCGAAGUCACAGAA 110% GC GAGG 14078 1041 2398 GCUAAAUUCUG 4302CACAGAAUUUAGCUC 99% UG GGUA 14079 1042 2399 CUAAAUUCUGU 4303CCACAGAAUUUAGCU 109% GG CGGU 14080 755 2400 AGACACGUUUG 4304GCCAAACGUGUCUUC 121% GC CAGU 14081 467 2401 CCGCAAGAUCG 4305GCCGAUCUUGCGGUU 132% GC GGCC 14082 995 2402 UAUCAAGUUU 4306CUCAAACUUGAUAGG 105% GAG CUUG 14083 927 2403 GAAGCUGACCU 4307CCAGGUCAGCUUCGC 114% GG AAGG 17356 1267 2404 ACAUUAACUCA 4308UAUGAGUUAAUGUC 120% UA UCUCA 17357 1267 2405 GACAUUAACUC 2406UAUGAGUUAAUGUC 56% AUA UCUCA 17358 1442 2407 UGAAGAAUGU 2408UUAACAUUCUUCAAA 34% UAA CCAG 17359 1442 2409 UUGAAGAAUG 2410UUAACAUUCUUCAAA 31% UUAA CCAG 17360 1557 2411 GAUAGCAUCUU 2412UUAAGAUGCUAUCU 59% AA GAUGA 17361 1557 2413 AGAUAGCAUCU 2414UUAAGAUGCUAUCU 47% UAA GAUGA 17362 1591 2415 UGAAGUGUAA 2416UAAUUACACUUCAAA 120% UUA UAGC 17363 1599 2417 AAUUGAGAAGG 2418UUCCUUCUCAAUUAC 71% AA ACUU 17364 1601 2419 UUGAGAAGGAA 2420UUUUCCUUCUCAAUU 62% AA ACAC 17365 1732 2421 CAUUCUGAUUC 2422UCGAAUCAGAAUGUC 99% GA AGAG 17366 1734 2423 UUCUGAUUCGA 2424UUUCGAAUCAGAAUG 97% AA UCAG 17367 1770 2425 CUGUCGAUUAG 2426UUCUAAUCGACAGGA 45% AA UUCC 17368 1805 2427 UUUGCCUGUAA 2428UGUUACAGGCAAAUU 71% CA CACU 17369 1805 2429 AUUUGCCUGUA 2430UGUUACAGGCAAAUU 67% ACA CACU 17370 1815 2431 ACAAGCCAGAU 2432UAAUCUGGCUUGUU 65% UA ACAGG 17371 1815 2433 AACAAGCCAGA 2434UAAUCUGGCUUGUU 35% UUA ACAGG 17372 2256 2435 CAGUUUAUUU 2436UACAAAUAAACUGUC 113% GUA CGAA 17373 2265 2437 UGUUGAGAGU 2438UACACUCUCAACAAA 35% GUA UAAA 17374 2265 2439 UUGUUGAGAG 2440UACACUCUCAACAAA 31% UGUA UAAA 17375 2295 2441 UGCACCUUUCU 2442UUAGAAAGGUGCAAA 34% AA CAUG 17376 2295 2443 UUGCACCUUUC 2444UUAGAAAGGUGCAAA 28% UAA CAUG 17377 1003 2445 UUGAGCUUUC 2446UCAGAAAGCUCAAAC 67% UGA UUGA 17378 2268 2447 UGAGAGUGUG 2448UGUCACACUCUCAAC 42% ACA AAAU 17379 2272 2449 AGUGUGACCAA 2450UUUUGGUCACACUCU 35% AA CAAC 17380 2272 2451 GAGUGUGACCA 2452UUUUGGUCACACUCU 29% AAA CAAC 17381 2273 2453 GUGUGACCAAA 2454UUUUUGGUCACACUC 42% AA UCAA 17382 2274 2455 UGUGACCAAAA 2456UCUUUUGGUCACACU 42% GA CUCA 17383 2274 2457 GUGUGACCAAA 2458UCUUUUGGUCACACU 37% AGA CUCA 17384 2275 2459 GUGACCAAAAG 2460UACUUUUGGUCACAC 24% UA UCUC 17385 2277 2461 GACCAAAAGUU 2462UUAACUUUUGGUCAC 27% AA ACUC 17386 2296 2463 GCACCUUUCUA 2464UCUAGAAAGGUGCAA 23% GA ACAU 17387 2299 2465 CCUUUCUAGUU 2466UCAACUAGAAAGGUG 46% GA CAAA

TABLE 16 Inhibition of gene expression with TGFB2 sd-rxRNA sequences(Accession Number: NM_001135599.1) % remaining Oligo Start SEQ ID SEQ IDexpression Number Site NO Sense sequence NO Antisense sequence (1 uM,A549) 14408 1324 2467 GGCUCUCCUUCGA 2468 UCGAAGGAGAGCCAU 94% UCGC 144091374 2469 GACAGGAACCUGG 2470 CCAGGUUCCUGUCUU n/a UAUG 14410 946 2471CCAAGGAGGUUUA 2472 UAAACCUCCUUGGCG 90% UAGU 14411 849 2473 AUUUCCAUCUACA2474 UGUAGAUGGAAAUCA 72% CCUC 14412 852 2475 UCCAUCUACAACA 2476UGUUGUAGAUGGAA 76% AUCAC 14413 850 2477 UUUCCAUCUACAA 2478UUGUAGAUGGAAAU 98% CACCU 14414 944 2479 CGCCAAGGAGGUU 2480AACCUCCUUGGCGUA 100% GUAC 14415 1513 2481 GUGGUGAUCAGAA 2482UUCUGAUCACCACUG n/a GUAU 14416 1572 2483 CUCCUGCUAAUGU 2484ACAUUAGCAGGAGAU 100% GUGG 14417 1497 2485 ACCUCCACAUAUA 2486UAUAUGUGGAGGUG 73% CCAUC 14418 1533 2487 AAGUCCACUAGGA 2488UCCUAGUGGACUUUA 98% UAGU 14419 1514 2489 UGGUGAUCAGAAA 2490UUUCUGAUCACCACU 86% GGUA 14420 1534 2491 AGUCCACUAGGAA 2492UUCCUAGUGGACUU 99% UAUAG 14421 943 2493 ACGCCAAGGAGGU 2494ACCUCCUUGGCGUAG 41% UACU 18570 2445 2495 UAUUUAUUGUGUA 2496UACACAAUAAAUAAC 79% UCAC 18571 2445 2497 UUAUUUAUUGUG 2498UACACAAUAAAUAAC 75% UA UCAC 18572 2083 2499 AUCAGUGUUAAAA 2500UUUUAACACUGAUGA 47% ACCA 18573 2083 2501 CAUCAGUGUUAAAA 2502UUUUAACACUGAUGA 17% ACCA 18574 2544 2503 AUGGCUUAAGGAA 2504UUCCUUAAGCCAUCC 59% AUGA 18575 2544 2505 GAUGGCUUAAGG 2506UUCCUUAAGCCAUCC 141% AA AUGA 18576 2137 2507 UUGUGUUCUGUUA 2508UAACAGAACACAAAC 77% UUCC 18577 2137 2509 UUUGUGUUCUGU 2510UAACAGAACACAAAC 59% UA UUCC 18578 2520 2511 AAAUACUUUGCCA 2512UGGCAAAGUAUUUG 75% GUCUC 18579 2520 2513 CAAAUACUUUGCCA 2514UGGCAAAGUAUUUG 55% GUCUC 18580 3183 2515 CUUGCACUACAAA 2516UUUGUAGUGCAAGU 84% CAAAC 18581 3183 2517 ACUUGCACUACAAA 2518UUUGUAGUGCAAGU 80% CAAAC 18582 2267 2519 GAAUUUAUUAGUA 2520UACUAAUAAAUUCUU 82% CCAG 18583 2267 2521 AGAAUUUAUUAG 2522UACUAAUAAAUUCUU 67% UA CCAG 18584 3184 2523 UUGCACUACAAAA 2524UUUUGUAGUGCAAG 77% UCAAA 18585 3184 2525 CUUGCACUACAAAA 2526UUUUGUAGUGCAAG 59% UCAAA 18586 2493 2527 AUAAAACAGGUGA 2528UCACCUGUUUUAUU 84% UUCCA 18587 2493 2529 AAUAAAACAGGUGA 2530UCACCUGUUUUAUU 70% UUCCA 18588 2297 2531 GACAACAACAACA 2532UGUUGUUGUUGUCG 40% UUGUU 18589 2046 2533 AUGCUUGUAACAA 2534UUGUUACAAGCAUCA 39% UCGU 18590 2531 2535 CAGAAACUCAUGA 2536UCAUGAGUUUCUGG 56% CAAAG 18591 2389 2537 GUAUUGCUAUGCA 2538UGCAUAGCAAUACAG 64% AAAA 18592 2530 2539 CCAGAAACUCAUA 2540UAUGAGUUUCUGGC 44% AAAGU 18593 2562 2541 ACUCAAACGAGCA 2542UGCUCGUUUGAGUU 87% CAAGU 18594 2623 2543 AUAUGACCGAGAA 2544UUCUCGGUCAUAUAA 69% UAAC 18595 2032 2545 CGACGACAACGAA 2546UUCGUUGUCGUCGU 55% CAUCA 18596 2809 2547 GUAAACCAGUGAA 2548UUCACUGGUUUACUA 58% AACU 18597 2798 2549 UUGUCAGUUUAGA 2550UCUAAACUGACAAAG 38% AACC 18598 2081 2551 UCAUCAGUGUUAA 2552UUAACACUGAUGAAC 25% CAAG 18599 2561 2553 AACUCAAACGAGA 2554UCUCGUUUGAGUUC 57% AAGUU 18600 2296 2555 CGACAACAACAAA 2556UUUGUUGUUGUCGU 69% UGUUC 18601 2034 2557 ACGACAACGAUGA 2558UCAUCGUUGUCGUCG 22% UCAU 18602 2681 2559 GCUGCCUAAGGAA 2560UUCCUUAGGCAGCUG 43% AUAC 18603 2190 2561 AUUCUACAUUUCA 2562UGAAAUGUAGAAUAA 128% GGCC

TABLE 17 Inhibition of gene expression with TGFB1 sd-rxRNA sequences(Accession Number NM_000660.3) % remaining Oligo Start SEQ ID SEQ IDexpression Number Site NO Sense sequence NO Antisense sequence (1 uMA549) 14394 1194 2563 GCUAAUGGUGGAA 2564 UUCCACCAUUAGCA 24% CGCGG 143952006 2565 UGAUCGUGCGCUC 2566 GAGCGCACGAUCAU 79% GUUGG 14396 1389 2567CAAUUCCUGGCGA 2568 UCGCCAGGAAUUGU 77% UGCUG 14397 1787 2569AGUGGAUCCACGA 2570 UCGUGGAUCCACUU n/a CCAGC 14398 1867 2571UACAGCAAGGUCC 2572 GGACCUUGCUGUAC 82% UGCGU 14399 2002 2573AACAUGAUCGUGC 2574 GCACGAUCAUGUUG n/a GACAG 14400 2003 2575ACAUGAUCGUGCG 2576 CGCACGAUCAUGUU n/a GGACA 14401 1869 2577CAGCAAGGUCCUG 2578 CAGGACCUUGCUGU 82% ACUGC 14402 2000 2579CCAACAUGAUCGU 2580 ACGAUCAUGUUGGA 66% CAGCU 14403 986 2581 AGCGGAAGCGCAU2582 AUGCGCUUCCGCUU 78% CACCA 14404 995 2583 GCAUCGAGGCCAU 2584AUGGCCUCGAUGCG 79% CUUCC 14405 963 2585 GACUAUCGACAUG 2586CAUGUCGAUAGUCU 80% UGCAG 14406 955 2587 ACCUGCAAGACUA 2588UAGUCUUGCAGGUG 88% GAUAG 14407 1721 2589 GCUCCACGGAGAA 2590UUCUCCGUGGAGCU n/a GAAGC 18454 1246 2591 CACAGCAUAUAUA 2592UAUAUAUGCUGUG 58% UGUACU 18455 1248 2593 CAGCAUAUAUAUA 2594UAUAUAUAUGCUGU 87% GUGUA 18456 1755 2595 GUACAUUGACUUA 2596UAAGUCAAUGUACA 107% GCUGC 18457 1755 2597 UGUACAUUGACUUA 2598UAAGUCAAUGUACA 77% GCUGC 18458 1708 2599 AACUAUUGCUUCA 2600UGAAGCAAUAGUUG 75% GUGUC 18459 1708 2601 CAACUAUUGCUUCA 2602UGAAGCAAUAGUUG 73% GUGUC 18460 1250 2603 GCAUAUAUAUGUA 2604UACAUAUAUAUGCU n/a GUGUG 18461 1754 2605 UGUACAUUGACUA 2606UAGUCAAUGUACAG 91% CUGCC 18462 1754 2607 CUGUACAUUGACUA 2608UAGUCAAUGUACAG 92% CUGCC 18463 1249 2609 AGCAUAUAUAUGA 2610UCAUAUAUAUGCUG n/a UGUGU 18464 1383 2611 CAGCAACAAUUCA 2612UGAAUUGUUGCUG 77% UAUUUC 18465 1251 2613 CAUAUAUAUGUUA 2614UAACAUAUAUAUGC 84% UGUGU 18466 1713 2615 UUGCUUCAGCUCA 2616UGAGCUGAAGCAAU n/a AGUUG 18467 1713 2617 AUUGCUUCAGCUCA 2618UGAGCUGAAGCAAU 83% AGUUG 18468 1247 2619 ACAGCAUAUAUAA 2620UUAUAUAUGCUGU 96% GUGUAC 18469 1712 2621 AUUGCUUCAGCUA 2622UAGCUGAAGCAAUA 90% GUUGG 18470 1712 2623 UAUUGCUUCAGCUA 2624UAGCUGAAGCAAUA 98% GUUGG 18471 1212 2625 CAAGUUCAAGCAA 2626UUGCUUGAACUUGU n/a CAUAG 18472 1222 2627 CAGAGUACACACA 2628UGUGUGUACUCUGC 45% UUGAA 18473 1228 2629 ACACACAGCAUAA 2630UUAUGCUGUGUGU 36% ACUCUG 18474 1233 2631 CAGCAUAUAUAUA 2632UAUAUAUAUGCUGU 68% GUGUA 18475 1218 2633 UCAAGCAGAGUAA 2634UUACUCUGCUUGAA 64% CUUGU 18476 1235 2635 AGCAUAUAUAUGA 2636UCAUAUAUAUGCUG 78% UGUGU 18477 1225 2637 AGAGUACACACAA 2638UUGUGUGUACUCU 92% GCUUGA 18478 1221 2639 AAGCAGAGUACAA 2640UUGUACUCUGCUUG 103% AACUU 18479 1244 2641 UUCAACACAUCAA 2642UUGAUGUGUUGAA 84% GAACAU 18480 1224 2643 AGCAGAGUACACA 2644UGUGUACUCUGCUU 37% GAACU 18481 1242 2645 AUAUAUGUUCUUA 2646UAAGAACAUAUAUA 62% UGCUG 18482 1213 2647 GACAAGUUCAAGA 2648UCUUGAACUUGUCA 47% UAGAU 18483 1760 2649 UUAAAGAUGGAGA 2650UCUCCAUCUUUAAU 69% GGGGC 18484 1211 2651 CUAUGACAAGUUA 2652UAACUUGUCAUAGA n/a UUUCG 19411 1212 2653 CAACGAAAUCUAA 2654UUAGAUUUCGUUG 52% UGGGUU 19412 1222 2655 UAUGACAAGUUCA 2656UGAACUUGUCAUAG 51% AUUUC 19413 1228 2657 AAGUUCAAGCAGA 2658UCUGCUUGAACUUG n/a UCAUA 19414 1233 2659 CAAGCAGAGUACA 2660UGUACUCUGCUUGA 41% ACUUG 19415 1218 2661 AAUCUAUGACAAA 2662UUUGUCAUAGAUU 104% UCGUUG 19416 1244 2663 CACACAGCAUAUA 2664UAUAUGCUGUGUG 31% UACUCU

TABLE 18 Inhibition of gene expression with SPP1 sd-rxRNA sequences(Accession Number NM_000582.2) % remaining Oligo Start SEQ ID SEQ IDexpression Number Site NO Sense sequence NO Antisense sequence (1 uMA549) 14084 1025 2665 mC.mU.mC. A.mU. 2666 P.mU.fC.fU. A. 61% G. A.A.mU.mU. A. A.fU.fU.fC. A.fU. G. A. G. A.Chl G* A* A* A*mU* A* C. 140851049 2667 mC.mU. G. A. G. 2668 P.mU. A. A.fU.fU. G. 50% G.mU.mC. A.A.fC.fC.fU.mC. A. G* A* A.mU.mU. A.Chl A* G* A*mU* G. 14086 1051 2669 G.A. G. G.mU.mC. 2670 P.mU.fU.fU. A. A.fU.fU. n/a A. A.mU.mU. A. A. G.A.fC.mC.mU.mC* A* A.Chl G* A* A* G* A. 14087 1048 2671 mU.mC.mU. G. A.G. 2672 P.mA. A.fU.fU. G. 69% G.mU.mC. A. A.fC.fC.fU.fC. A. G. A*A.mU.mU.Chl A* G* A*mU* G* C. 14088 1050 2673 mU. G. A. G. 2674 P.mU.fU.A. A.fU.fU. G. 76% G.mU.mC. A. A.fC.fC.mU.mC. A* G* A.mU.mU. A. A.Chl A*A* G* A* U. 14089 1047 2675 mU.mU.mC.mU. G. 2676 P.mA.fU.fU. G. 60% A.G. G.mU.mC. A. A.fC.fC.fU.fC. A. G. A. A.mU.Chl A* G* A*mU* G*mC* A.14090 800 2677 G.mU.mC. A. 2678 P.mU.fC. A.fU.fC.fC. A. 71% G.mC.mU. G.G. G.fC.fU. G. A.mU. G. A.Chl A.mC*mU*mC* G*mU*mU* U. 14091 492 2679mU.mU.mC.mU. G. 2680 P.mA. G. A.fU.fU.fC. n/a A.mU. G. A. A.fU.fC. A. G.A. A*mU* A.mU.mC.mU.Chl G* G*mU* G* A. 14092 612 2681 mU. G. G. A.mC.mU.2682 P.mU. G. A.fC.fC.fU.fC. n/a G. A. G. G.mU.mC. A. G.fU.mC.mC. A*mU*A.Chl A* A* A*mC* C. 14093 481 2683 G. A. 2684 P.mA. A.fU. G. G.fU. G.n/a G.mU.mC.mU.mC. A. G. A.mC.mU.mC* A.mC.mC. A*mU*mC* A* G* A.A.mU.mU.Chl 14094 614 2685 G. A.mC.mU. G. A. 2686 P.mU.fU.fU. G. n/a G.G.mU.mC. A. A. A.fC.fC.fU.fC. A. A.Chl G.mU.mC*mC* A*mU* A* A* A. 14095951 2687 mU.mC. A.mC. A. 2688 P.mU.fU.fC. A.fU. G. 89% G.mC.mC. A.mU. G.G.fC.fU. G.mU. G. A* A* A. A.Chl A*mU*mU*mC* A. 14096 482 2689 A. 2690P.mG. A. A.fU. G. G.fU. 87% G.mU.mC.mU.mC. G. A. G. A.mC.mU*mC* A.mC.mC.A*mU*mC* A* G. A.mU.mU.mC.Chl 14097 856 2691 A. A. G.mC. G. G. A. 2692P.mU. G. 88% A. A. G.mC.mC. A.Chl G.fC.fU.fU.fU.fC.fC. G.mC.mU.mU* A*mU*A*mU* A* A. 14098 857 2693 A. G.mC. G. G. A. A. 2694 P.mU.fU. G. 113% A.G.mC.mC. A. A.Chl G.fC.fU.fU.fU.fC.fC. G.mC.mU*mU* A*mU* A*mU* A. 14099365 2695 A.mC.mC. A.mC. 2696 P.mU.fC. A.fU.fC.fC. 98% A.mU. G. G. A.mU.A.fU. G.fU. G. G. A.Chl G.mU*mC* A*mU* G* G* C. 14100 359 2697 G.mC.mC.A.mU. G. 2698 P.mA.fU. G.fU. G. 84% A.mC.mC. A.mC. G.fU.fC. A.fU. G.A.mU.Chl G.mC*mU*mU*mU*mC* G* U. 14101 357 2699 A. A. G.mC.mC. 2700P.mG.fU. G. G.fU.fC. 88% A.mU. G. A.mC.mC. A.fU. G. A.mC.ChlG.mC.mU.mU*mU*mC* G*mU*mU* G. 14102 858 2701 G.mC. G. G. A. A. A. 2702P.mA.fU.fU. G. n/a G.mC.mC. A. G.fC.fU.fU.fU.fC.mC. A.mU.Chl G.mC*mU*mU*A*mU* A* U. 14103 1012 2703 A. A. 2704 P.mA. A. A.fU. A.fC. G. 93%A.mU.mU.mU.mC. A. A. G.mU. A.mU.mU.mU*mC* A* A.mU.mU.mU.Chl G* G*mU* G.14104 1014 2705 A.mU.mU.mU.mC. 2706 P.mA. G. A. A. A.fU. 89% G.mU. A.fC.G. A. A. A.mU.mU.mU.mC.mU. A.mU*mU*mU*mC* Chl A* G* G. 14105 356 2707 A.A. A. G.mC.mC. 2708 P.mU. G. G.fU.fC. A.fU. 85% A.mU. G. A.mC.mC. G.A.Chl G.fC.mU.mU.mU*mC* G*mU*mU* G* G. 14106 368 2709 A.mC. A.mU. G. G.2710 P.mA.fU. A.fU.fC. 67% A.mU. G. A.mU. A.fU.fC.fC. A.mU. A.mU.ChlG.mU* G* G*mU*mC* A* U. 14107 1011 2711 G. A. A. 2712 P.mA. A.fU. A.fC.G. A. 87% A.mU.mU.mU.mC. A. A.fU.mU.mU.mC* A* G.mU. A.mU.mU.Chl G* G*mU*G* U. 14108 754 2713 G.mC. 2714 P.mA. A.fU.fC. A. G. A. 73%G.mC.mC.mU.mU.mC. A. G. G.mC. G.mC* mU. G. G*mU*mU*mC* A* G. A.mU.mU.Chl14109 1021 2715 A.mU.mU.mU.mC.mU. 2716 P.mA.fU.fU.fC. A.fU. G. 128% mC.A.mU. G. A. A. G. A. A. A.mU* A.mU.Chl A*mC* G* A* A* A. 14110 1330 2717mC.mU.mC.mU.mC. 2718 P.mC.fU. A.fU.fU.fC. 101% A.mU. G. A. A.mU. A.A.fU. G. A. G. A. G* A* G.Chl A*mU* A* A* C. 14111 346 2719 A. A.G.mU.mC.mC. 2720 P.mU.fU.fU.fC. G.fU.fU. 59% A. A.mC. G. A. A. G. G.A.mC.mU.mU* A.Chl A*mC*mU*mU* G* G. 14112 869 2721 A.mU. G. A.mU. G.2722 P.mU.fU. 89% A. G. A. G.mC. A. G.fC.fU.fC.fU.fC. A.Chl A.fU.mC.A.mU*mU* G* G*mC*mU* U. 14113 701 2723 G.mC. G. A. G. G. A. 2724P.mU.fU.fC. A. 95% G.mU.mU. G. A. A.fC.fU.fC.fC.fU.mC. A.ChlG.mC*mU*mU*mU*mC* mC* A. 14114 896 2725 mU. G. A.mU.mU. G. 2726 P.mU. G.A.fC.fU. 87% A.mU. A. G.mU.mC. A.fU.fC. A. A.mU.mC. A.Chl A*mC* A*mU*mC*G* G. 14115 1035 2727 A. G. A.mU. A. 2728 P.mA. G. A.fU. G.fC. 82% G.mU.G.mC. A.fC.fU. A.mU.mC.mU* A.mU.mC.mU.Chl A* A*mU*mU*mC* A. 14116 11702729 A.mU. G.mU. G.mU. 2730 P.mA. A.fU. A. G. A.fU. 36% A.mU.mC.mU.A.fC. A.mC. A.mU.mU.Chl A.mU*mU*mC* A* A*mC* C. 14117 1282 2731mU.mU.mC.mU. 2732 P.mU.fU.fC.fU.fU.fC.fU. 91% A.mU. A. G. A. A. G. A.fU.A. G. A. A*mU* A. A.Chl G* A* A*mC* A. 14118 1537 2733 mU.mU. 2734 P.mA.A.fU.fU. G.fC.fU. 152% G.mU.mC.mC. A. G. G. A.mC. A. G.mC. A. A*mC*mC*G*mU* G* A.mU.mU.Chl G. 14119 692 2735 A.mC. A.mU. G. G. 2736 P.mU.fC.n/a A. A. A. G. C.mG. G.fC.fU.fU.fU.fC.fC. A.Chl A.mU. G.mU* G*mU* G* A*G* G. 14120 840 2737 G.mC. A. 2738 P.mU. A. A.fU.fC.fU. G. 87%G.mU.mC.mC. A. G. G. A.fC.mU. A.mU.mU. A.Chl G.mC*mU*mU* G*mU* G* G.14121 1163 2739 mU. G. G.mU.mU. G. 2740 P.mA.fC. A.fC. 31% A. A.mU.G.mU. A.fU.fU.fC. A. A.mC.mC. G.mU.Chl A* A*mU* A* A* A* C. 14122 7892741 mU.mU. A.mU. G. A. 2742 P.mA.fC.fU.fC. 96% A. A.mC. G. A.G.fU.fU.fU.fC. A.mU. A. G.mU.Chl A*mC*mU* G*mU*mC* C. 14123 841 2743 mC.A. 2744 P.mA.fU. A. A.fU.fC.fU. 110% G.mU.mC.mC. A. G. G. G. A.mC.mU.A.mU.mU. A.mU.Chl G*mC*mU*mU* G*mU* G. 14124 852 2745 A.mU. A.mU. A. A.2746 P.mU.fU.fU.fC.fC. 91% G.mC. G. G. A. A. G.fC.fU.fU. A.mU. A.ChlA.mU* A* A*mU*mC*mU* G. 14125 209 2747 mU. A.mC.mC. A. 2748 P.mU.G.fU.fU.fU. A. 110% G.mU.mU. A. A. A.fC.fU. G. G.mU. A.mC. A.Chl A*mU*G* G*mC* A* C. 14126 1276 2749 mU. G.mU.mU.mC. 2750 P.mU. A.fU. A. G. A.n/a A.mU.mU.mC.mU. A.fU. G. A. A.mC. A.mU. A.Chl A*mU* A* G* A*mC* A.14127 137 2751 mC.mC. G. A.mC.mC. 2752 P.mU.fU.fU.fC.fC.fU.fU. 71% A. A.G. G. A. A. G. G.fU.mC. G. G*mC* A.Chl G*mU*mU*mU* G. 14128 711 2753 G.A. A.mU. G. 2754 P.mG.fU. A.fU. G.fC. 115% G.mU. G.mC. A.mU. A.fC.fC.A.mU.mU.mC* A.mC.Chl A* A*mC*mU*mC* C. 14129 582 2755 A.mU. A.mU. G.2756 P.mU.fC. G. G.fC.fC. 97% A.mU. G. G.mC.mC. A.fU.fC. A.mU. A.mU* G.A.Chl G*mU* G*mU*mC* U. 14130 839 2757 A. G.mC. A. 2758 P.mA.A.fU.fC.fU. G. G. 102% G.mU.mC.mC. A. G. A.fC.fU. G.mC.mU*mU*A.mU.mU.Chl G*mU* G* G* C. 14131 1091 2759 G.mC. 2760 P.mU.fU.fU. G.A.fC.fU. 10% A.mU.mU.mU. A. A. A. A.mU. G.mC* A* G.mU.mC. A. A. A* A*G*mU* G. A.Chl 14132 884 2761 A. G.mC. 2762 P.mA.fC. A.fU.fC. G. G. 93%A.mU.mU.mC.mC. G. A. A.fU. G.mC.mU*mC* A.mU. G.mU.Chl A*mU*mU* G* C.14133 903 2763 mU. A. G.mU.mC. A. 2764 P.mA. A. 97% G. G. A.G.fU.fU.fC.fC.fU. G. A.mC.mU.mU.Chl A.mC.mU. A*mU*mC* A* A*mU* C. 141341090 2765 mU. G.mC. 2766 P.mU.fU. G. A.fC.fU. A. 39% A.mU.mU.mU. A. A.A.fU. G.mC. A* A* A* G.mU.mC. A. A.Chl G*mU* G* A. 14135 474 2767G.mU.mC.mU. G. 2768 P.mA. G. A.fC.fU.fC. 99% A.mU. G. A. A.fU.fC. A. G.G.mU.mC.mU.Chl A.mC*mU* G* G*mU* G* A. 14136 575 2769 mU. A. G. A.mC.2770 P.mU.fC. A.fU. A.fU. 108% A.mC. A.mU. A.mU. G.fU. G.fU.mC.mU. G.A.Chl A*mC*mU* G*mU* G* G. 14137 671 2771 mC. A. G. A.mC. G. 2772P.mA.fU. 98% A. G. G. A.mC. G.fU.fC.fC.fU.fC. A.mU.Chl G.fU.mC.mU. G*mU*A* G*mC* A* U. 14138 924 2773 mC. A. G.mC.mC. 2774 P.mG. A. A.fU.fU.fC.100% G.mU. G. A. A.fC. G. G.mC.mU. G* A.mU.mU.mC.Chl A*mC*mU*mU*mU* G.14139 1185 2775 A. G.mU.mC.mU. G. 2776 P.mU.fU. 47% G. A. A. A.mU. A.A.fU.fU.fU.fC.fC. A. G. A.Chl A.mC.mU*mC* A* A* A*mU* A. 14140 1221 2777A. G.mU.mU.mU. 2778 P.mG. A. A. G.fC.fC. 100% G.mU. G. A.fC. A. A.A.mC.mU* G.mC.mU.mU.mC.Chl A* A* A*mC*mU* A. 14141 347 2779 A.G.mU.mC.mC. A. 2780 P.mC.fU.fU.fU.fC. 103% A.mC. G. A. A. A. G.fU.fU. G.G. G.Chl A.mC.mU*mU* A*mC*mU*mU* G. 14142 634 2781 A. A. 2782P.mG.fU.fC.fU. G.fC. G. 100% G.mU.mU.mU.mC. A. A. G.mC. A. G.A.mC.mU.mU*mC*mU* A.mC.Chl mU* A* G* A. 14143 877 2783 A. G.mC. A. A.mU.2784 P.mA. A.fU. G.fC.fU.fC. 104% G. A. G.mC. A.fU.fU. A.mU.mU.ChlG.mC.mU*mC*mU*mC* A*mU* C. 14144 1033 2785 mU.mU. A. G. A.mU. 2786P.mA.fU. G.fC. A.fC.fU. 95% A. G.mU. G.mC. A.fU.fC.mU. A. A.mU.ChlA*mU*mU*mC* A*mU* G. 14145 714 2787 mU. G. G.mU. G.mC. 2788 P.mC.fU.fU.G.fU. A.fU. 101% A.mU. A.mC. A. A. G.fC. A.mC.mC. G.Chl A*mU*mU*mC* A*A* C. 14146 791 2789 A.mU. G. A. A. 2790 P.mU. G. A.fC.fU.fC. 100% A.mC.G. A. G.fU.fU.fU.mC. A.mU* G.mU.mC. A.Chl A* A*mC*mU* G* U. 14147 8132791 mC.mC. A. G. A. 2792 P.mU.fU.fC. A. G.fC. 97% G.mU. G.mC.mU. G.A.fC.fU.fC.mU. G. A. A.Chl G*mU*mC* A*mU*mC* C. 14148 939 2793 mC. A.G.mC.mC. 2794 P.mA. A. A.fU.fU.fC. 109% A.mU. G. A. A.fU. G. G.mC.mU.A.mU.mU.mU.Chl G*mU* G* G* A* A* U. 14149 1161 2795 A.mU.mU. G. 2796P.mA.fC. A.fU.fU.fC. A. 34% G.mU.mU. G. A. A.fC.fC. A. A.mU* A* A* A.mU.G.mU.Chl A*mC*mU* G. 14150 1164 2797 G. G.mU.mU. G. A. 2798 P.m U. A.fC.A.fC. n/a A.mU. G.mU. G.mU. A.fU.fU.fC. A. A.Chl A.mC.mC* A* A*mU* A* A*A. 14151 1190 2799 G. G. A. A. A.mU. A. 2800 P.mA.fU.fU. A. G.fU.fU. n/aA.mC.mU. A. A.fU.fU.mU.mC.mC* A* A.mU.Chl G* A*mC*mU* C. 14152 1333 2801mU.mC. A.mU. G. A. 2802 P.mU.fU.fU.fC.fU. 31% A.mU. A. G. A. A.A.fU.fU.fC. A.mU. G. A* A.Chl G* A* G* A* A* U. 14153 537 2803 G.mC.mC.A. G.mC. 2804 P.mU.fU.fC. G. G.fU.fU. n/a A. A.mC.mC. G. A. G.fC.fU. G.G.mC* A* A.Chl G* G*mU*mC* C. 14154 684 2805 mC. 2806 P.mC. A.fU. G.fU.G.fU. 100% A.mC.mC.mU.mC. G. A. G. G.mU. G* A.mC. A.mC. A.mU. A*mU*G*mU*mC* C. G.Chl 14155 707 2807 A. G.mU.mU. G. A. 2808 P.mG.fC.A.fC.fC. 99% A.mU. G. G.mU. A.fU.fU.fC. A. G.mC.Chl A.mC.mU*mC*mC*mU*mC* G* C. 14156 799 2809 A. G.mU.mC. A. 2810 P.mC. A.fU.fC.fC. A. 95%G.mC.mU. G. G. G.fC.fU. G. A.mU. G.Chl A.mC.mU*mC* G*mU*mU*mU* C. 14157853 2811 mU. A.mU. A. A. 2812 P.mC.fU.fU.fU.fC.fC. 106% G.mC. G. G. A.A. A. G.fC.fU.fU. A.mU. G.Chl A*mU* A* A*mU*mC* U. 14158 888 2813mU.mU.mC.mC. G. 2814 P.mA. A.fU.fC. A.fC. 88% A.mU. G.mU. G. A.fU.fC. G.G. A. A*mU* A.mU.mU.Chl G*mC*mU*mC* A. 14159 1194 2815 A.mU. A. A.mC.mU.2816 P.mA.fC. A.fC. A.fU.fU. 95% A. A.mU. G.mU. A. G.fU.mU. G.mU.ChlA.mU*mU*mU*mC*mC* A* G. 14160 1279 2817 mU.mC. 2818 P.mU.fU.fC.fU. A.fU.A. 15% A.mU.mU.mC.mU. G. A. A.mU. G. A* A.mU. A. G. A. A.Chl A*mC* A*mU*A*G. 14161 1300 2819 A. A.mC.mU. 2820 P.mU. A.fC. A. G.fU. G. 86%A.mU.mC. A.mC.mU. A.fU. A. G.mU. A.Chl G.mU.mU*mU* G*mC* A*mU* U. 141621510 2821 G.mU.mC. A. 2822 P.mA.fU. A. A. G.fC. A. 86% A.mU.mU. A.fU.fU.G. A.mC* G.mC.mU.mU. A*mC*mC* A*mC* C. A.mU.Chl 14163 1543 2823 A. G.mC.A. 2824 P.mU.fU.fU. A.fU.fU. A. 110% A.mU.mU. A. A.mU. A.fU.fU. G.mC.mU*G* A. A. A.Chl G* A*mC* A* A. 14164 434 2825 A.mC. G. 2826 P.mU.fC.A.fU.fC. A. G. 134% A.mC.mU.mC.mU. G. A. G.fU.mC. A.mU. G. A.ChlG.mU*mU*mC* G* A* G* U. 14165 600 2827 mU. A. G.mU. G.mU. 2828 P.mA.fU.A. A. A.fC.fC. 102% G. G.mU.mU.mU. A.fC. A.mC.mU. A.mU.Chl A*mU*mC*A*mC*mC* U. 14166 863 2829 A. A. G.mC.mC. A. 2830 P.mU.fC. A.fU.fC. 93%A.mU. G. A.mU. G. A.fU.fU. G. A.Chl G.mC.mU.mU*mU*mC* mC* G*mC* U. 14167902 2831 A.mU. A. G.mU.mC. 2832 P.mA. G.fU.fU.fC.fC.fU. 101% A. G. G. A.G.A.fC.mU. A.mU*mC* A.mC.mU.Chl A* A*mU*mC* A. 14168 921 2833 A.G.mU.mC. A. 2834 P.mU.fU.fC. A.fC. G. 98% G.mC.mC. G.mU. G. G.fC.fU. G.A. A.Chl A.mC.mU*mU*mU* G* G* A* A. 14169 154 2835 A.mC.mU. 2836P.mU.fU.fC.fU.fC. A.fU. n/a A.mC.mC. A.mU. G. G. G.fU. A. G.mU* G* A. G.A. A.Chl A* G*mU*mU* U. 14170 217 2837 A. A. A.mC. A. G. 2838 P.mA.A.fU.fC. A. 66% G.mC.mU. G. G.fC.fC.fU. A.mU.mU.Chl G.mU.mU.mU* A*A*mC*mU* G* G. 14171 816 2839 G. A. G.mU. 2840 P.mG. G.fU.fU.fU.fC. A.102% G.mC.mU. G. A. A. G.fC. A.mC.mC.Chl A.mC.mU.mC*mU* G* G*mU*mC* A.14172 882 2841 mU. G. A. G.mC. 2842 P.mA.fU.fC. G. G. A. 103%A.mU.mU.mC.mC. G. A.fU. G.fC.mU.mC. A.mU.Chl A*mU*mU* G*mC*mU* C. 14173932 2843 A. 2844 P.mU. G. G.fC.fU. G.fU. n/a A.mU.mU.mC.mC. G. G. A.A.mU.mU*mC* A.mC. A. G.mC.mC. A*mC* G* G* C. A.Chl 14174 1509 2845 mU.G.mU.mC. A. 2846 P.mU. A. A. G.fC. A. n/a A.mU.mU. A.fU.fU. G. A.mC.G.mC.mU.mU. A.Chl A*mC*mC* A*mC*mC* A. 14175 157 2847 A.mC.mC. A.mU. G.2848 P.mC. A. 109% A. G. A. A.mU.mU. A.fU.fU.fC.fU.fC. A.fU. G.Chl G.G.mU* A* G*mU* G* A* G. 14176 350 2849 mC.mC. A. A.mC. G. 2850 P.mU. G.95% A. A. A. G.mC.mC. G.fC.fU.fU.fU.fC. A.Chl G.fU.mU. G. G* A*mC*mU*mU*A* C. 14177 511 2851 mC.mU. G. 2852 P.mA. A.fU.fC. A. G.fU. 100%G.mU.mC. A.mC.mU. G. A.fC.mC. A. G. A.mU.mU.Chl G*mU*mU*mC* A*mU* C.14178 605 2853 mU. G. 2854 P.mA. G.fU.fC.fC. A.fU. 99% G.mU.mU.mU. A. A.A.mC.mC. A*mC* A.mU. G. G. A*mC*mU* A* U. A.mC.mU.Chl 14179 811 2855 G.A.mC.mC. A. G. 2856 P.mC. A. G.fC. 88% A. G.mU. G.mC.mU. A.fC.fU.fC.fU.G. G.Chl G.mU.mC* A*mU*mC*mC* A* G. 14180 892 2857 G. A.mU. G.mU. G.2858 P.mU. A.fU.fC. A. 76% A.mU.mU. G. A.mU. A.fU.fC. A.fC. A.ChlA.mU.mC* G* G* A* A*mU* G. 14181 922 2859 G.mU.mC. A. 2860P.mA.fU.fU.fC. A.fC. G. 59% G.mC.mC. G.mU. G. G.fC.fU. G. A. A.mU.ChlA.mC*mU*mU*mU* G* G* A. 14182 1169 2861 A. A.mU. G.mU. 2862 P.mA.fU. A.G. A.fU. 69% G.mU. A.fC. A.fC. A.mU.mC.mU. A.mU.mU*mC* A* A.mU.ChlA*mC*mC* A. 14183 1182 2863 mU.mU. G. A. 2864 P.mU.fU.fU.fC.fC. A. G.n/a G.mU.mC.mU. G. G. A.fC.fU.mC. A. A* A. A. A.Chl A*mU* A* G* A* U.14184 1539 2865 G.mU.mC.mC. A. 2866 P.mU.fU. A. A.fU.fU. 77% G.mC. A.A.mU.mU. G.fC.fU. G. G. A.mC* A* A. A.Chl A*mC*mC* G* U. 14185 1541 2867mC.mC. A. G.mC. A. 2868 P.mU. A.fU.fU. A. n/a A.mU.mU. A. A.mU. A.fU.fU.G.fC.mU. G. G* A.Chl A*mC* A* A*mC*C. 14186 427 2869 G. A.mC.mU.mC. G.2870 P.mA. G.fU.fC. 69% A. A.mC. G. G.fU.fU.fC. G. A. A.mC.mU.ChlG.mU.mC* A* A*mU* G* G* A. 14187 533 2871 A.mC.mC.mU. 2872 P.mG.fU.fU.G.fC.fU. G. 78% G.mC.mC. A. G.mC. G.fC. A. G. A. A.mC.Chl G.mU*mC*mC*G*mU* G* G. 18538 496 2873 G. A.mU. G. A. 2874 P.mU. A.fU.fC. A. G. 74%A.mU.mC.mU. G. A.fU.fU.fC. A.fU.fC* A* A.mU. A.Chl G* A* A*fU* G. 18539496 2875 mU. G. A.mU. G. A. 2876 P.mU. A.fU.fC. A. G. 72% A.mU.mC.mU. G.A.fU.fU.fC. A.fU.fC* A* A.mU. A.Chl G* A* A*fU* G. 18540 175 2877A.mU.mU.mU. 2878 P.mU. G.fC. A. A. A. A. 98% G.mC.mU.mU.mU.mU. G.fC. A.A. A.fU*fC* G.mC. A.Chl A*fC*fU*fG* C. 18541 175 2879 G. A.mU.mU.mU.2880 P.mU. G.fC. A. A. A. A. 28% G.mC.mU.mU.mU.mU. G.fC. A. A. A.fU*fC*G.mC. A.Chl A*fC*fU*fG* C. 18542 172 2881 G.mU. G. 2882 P.mU. A. A. A.G.fC. A. 24% A.mU.mU.mU. A. A.fU.fC. A.fC*fU* G.mC.mU.mU.mU. G*fC* A* A*U. A.Chl 18543 172 2883 A. G.mU. G. 2884 P.mU. A. A. A. G.fC. A. 14%A.mU.mU.mU. A. A.fU.fC. A.fC*fU* G.mC.mU.mU.mU. G*fC* A* A* U. A.Chl18544 1013 2885 A. 2886 P.mU. A. A. A.fU. A.fC. 100% A.mU.mU.mU.mC. G.A. A. A.fU.fU*fU*fC* G.mU. A* G* G* U. A.mU.mU.mU. A.Chl 18545 1013 2887A. A. 2888 P.mU. A. A. A.fU. A.fC. 109% A.mU.mU.mU.mC. G. A. A.A.fU.fU*fU*fC* G.mU. A* G* G* U. A.mU.mU.mU. A.Chl 18546 952 2889 mC.A.mC. A. 2890 P.mU.fU.fU. C. A.fU. G. 32% G.mC.mC. A.mU. G. G.fC.fU.G.fU. G* A* A* A. A. A.Chl A*fU*fU* C. 18547 952 2891 mU.mC. A.mC. A.2892 P.mU.fU.fU. C. A.fU. G. 33% G.mC.mC. A.mU. G. G.fC.fU. G.fU. G* A*A* A. A. A.Chl A*fU*fU* C. 18548 174 2893 G. A.mU.mU.mU. 2894 P.mU.fC.A. A. A. A. 57% G.mC.mU.mU.mU.mU. G.fC. A. A. A.fU.fC* G. A.Chl A*fC*fU*G*fC* A. 18549 174 2895 mU. G. 2896 P.mU.fC. A. A. A. A. 53% A.mU.mU.mU.G.fC. A. A. A.fU.fC* G.mC.mU.mU.mU.mU. A*fC*fU* G*fC* A. G. A.Chl 18550177 2897 mU.mU. 2898 P.mU. A. G. G.fC. A. A. 97% G.mC.mU.mU.mU.mU. A. A.G.fC. A. A* G.mC.mC.mU. A*fU*fC* A*fC* U. A.Chl 18551 177 2899 mU.mU.mU.2900 P.mU. A. G. G.fC. A. A. 103% G.mC.mU.mU.mU.mU. A. A. G.fC. A. A*G.mC.mC.mU. A*fU*fC* A*fC* U. A.Chl 18552 1150 2901 mU.mU.mU.mC.mU. 2902P.mU.fU. A. A. A.fC.fU. 96% mC. A. G. A. G. A. A. A* G* A* G.mU.mU.mU.A. A* G*fC* A. A.Chl 18553 1089 2903 mU.mU. G.mC. 2904 P.mU. G. A.fC.fU.A. A. 94% A.mU.mU.mU. A. A.fU. G.fC. A. A* A* G.mU.mC. A.Chl G*fU* G* A*G. 18554 1086 2905 A.mC.mU.mU.mU. 2906 P.mU.fU. A. A. A.fU. n/a G.mC.G.fC. A. A. A. G.fU* G* A.mU.mU.mU. A. A* G* A* A* A. A.Chl 18555 10932907 A.mU.mU.mU. A. 2908 P.mU.fU.fU.fU.fU. G. n/a G.mU.mC. A. A. A. A.A.fC.fU. A. A. A.fU* A.Chl G*fC* A* A* A* G. 18556 1147 2909mU.mU.mC.mU.mU. 2910 P.mU. A.fC.fU. G. A. G. n/a mU.mC.mU.mC. A. A. A.A. G. A. A* G*fC* G.mU. A.Chl A*fU*fU* U. 18557 1148 2911mU.mC.mU.mU.mU. 2912 P.mU. A. A.fC.fU. G. A. 66% mC.mU.mC. A. G. A. A.A. G. A* A* G.mU.mU. A.Chl G*fC* A*fU* U. 18558 1128 2913 G. A. A. A. G.A. G. 2914 P.mU. A.fU. 16% A. A.mC. A.mU. G.fU.fU.fC.fU.fC.fU.fU.fU.A.Chl fC* A*fU*fU*fU*fU* G. 18559 1087 2915 mC.mU.mU.mU. 2916P.mU.fC.fU. A. A. A.fU. 28% G.mC. G.fC. A. A. A. G*fU* G* A.mU.mU.mU. A.G. A* G* A* A. A.Chl 18560 1088 2917 mU.mU.mU. G.mC. 2918 P.mU. A.fC.fU.A. A. n/a A.mU.mU.mU. A. A.fU. G.fC. A. A. A* G.mU. A.Chl G*fU* G* A* G*A. 18561 1083 2919 mC.mU.mC. 2920 P.mU. A.fU. G.fC. A. A. 53%A.mC.mU.mU.mU. A. G.fU. G. A. G* A* A* G.mC. A.mU. A.Chl A*fU*fU* G.18562 1081 2921 mU.mU.mC.mU.mC. 2922 P.mU. G.fC. A. A. A. 89%A.mC.mU.mU.mU. G.fU. G. A. G. A. A* G.mC. A.Chl A*fU*fU* G*fU* A. 18563555 2923 mC. 2924 P.mU. A.fC. A. A.fC.fU. 33% A.mC.mU.mC.mC. A. G. G. A.G.fU. G* A* A* G.mU.mU. G.mU. A* A*fC*fU. A.Chl 18564 1125 2925 A. A.mU.G. A. A. A. 2926 P.mU.fU.fU.fC.fU.fC.fU. n/a G. A. G. A. A. A. ChlfU.fU.fC. A.fU.fU*fU*fU* G*fC*fU* A. 18565 168 2927 mU. G.mC. A. G.mU.2928 P.mU.fC. A. A. A.fU.fC. 14% G. A.fC.fU. G.fC. A* A.mU.mU.mU.mG.A*fU*fU*fC*fU* C. A.Chl 18566 1127 2929 mU. G. A. A. A. G. A. 2930P.mU.fU. 27% G. A. A.mC. A. A.Chl G.fU.fU.fC.fU.fC.fU.fU.fU. fC.A*fU*fU*fU*fU* G* C. 18567 1007 2931 A.mC.mC.mU. G. A. 2932 P.mU. G. A.A. 129% A. A.fU.fU.fU.fC. A. G. A.mU.mU.mU.mC. G.fU* G*fU*fU*fU* A*A.Chl U. 18568 164 2933 G. A. A.mU.mU. 2934 P.mU.fU.fC. A.fC.fU. 47%G.mC. A. G.mU. G. A. G.fC. A. A.Chl A.fU.fU.fC*fU*fC* A*fU* G* G. 18569222 2935 G. G.mC.mU. G. 2936 P.mU.fC.fC. A. G. A. n/a A.mU.mU.mC.mU.A.fU.fC. A. G.fC.fC*fU* G. G. A.Chl G*fU*fU*fU* A. 20612 172 2937 A.G.mU. G. 2938 P.mU. A. A. A. G.fC. A. n/a A.mU.mU.mU. A. A.fU.mC.A.mC*mU* G.mC.mU.mU.mU. G*mC* A* A* U. A.Chl 20613 172 2939 A. G.mU. G.2940 P.mU. A. A. A. G.fC. A. n/a A.mU.mU.mU. A. A.fU.fC. A.mC*fU*G.mC.mU.mU.mU. G*mC* A* A* U. A.Chl 20614 172 2941 A. G.mU. G. 2942P.mU. A. A. A. G. C. A. 101% A.mU.mU.mU. A. A. U.mC. A.mC*mU*G.mC.mU.mU.mU. G*mC* A* A* U. A.Chl 20615 172 2943 A. G.mU. G. 2944P.mU. A. A. A. G.fC. A. 104% A.mU.mU.mU. A. A.fU.mC. G.mC.mU.mU.mU.A.mC*mU*mG*mC*mA* A.Chl mA* U.

TABLE 19 Inhibition of gene expression with PTGS2 sd-rxRNA sequences(Accession Number: NM_000963.2) Oligo Start SEQ ID SEQ ID Number Site NOSense sequence NO Antisense sequence % remaining expression (1 uM A549)14422 451 2945 mC. A.mC. 2946 P.mU.fC. A. A.fU.fC. A. 72% A.mU.mU.mU. G.A. A.fU. G.mU. G* A.mU.mU. G. A.Chl A*mU*mC*mU* G* G. 14423 1769 2947mC. A.mC.mU. 2948 P.mA. A.fU.fU. G.A. G. 71% G.mC.mC.mU.mC. A. G.fC. A.G.mU. A.mU.mU.Chl G*mU*mU* G* A*mU* G. 14424 1464 2949 A. A. A.mU. 2950P.mA. A. G. A.fC.fU. G. 74% A.mC.mC. A. G.fU. G.mU.mC.mU.mU.ChlA.mU.mU.mU*mC* A*mU*mC*mU* G. 14425 453 2951 mC. A.mU.mU.mU. 2952 P.mU.G.fU.fC. A. 83% G. A.mU.mU. G. A.fU.fC. A. A. A.mU. A.mC. A.Chl G*mU* G*A*mU*mC* U. % remaining expression (1 uM PC-3) 17388 285 2953 G. A. A.A. 2954 P.mU.fU. G. A. G.fC. A. 88% A.mC.mU. G.fU.fU.fU.fU.fC*fU*fC*G.mC.mU.mC. A. fC* A*fU* A. A.Chl 17389 520 2955 A.mC.mC.mU.mC.mU. 2956P.mU. A. A.fU. A. G. G. 25% mC.mC.mU. A. G. A. G. G.fU*fU* A* A.mU.mU.A.Chl G* A* G* A. 17390 467 2957 mU.mC.mC. 2958 P.mU.fU. A. A. G.fU.fU.68% A.mC.mC. A. G. G.fU. G. G. A*fC*fU* A.mC.mU.mU. A. G*fU*fC* A. A.Chl17391 467 2959 G.mU.mC.mC. 2960 P.mU.fU. A. A. G.fU.fU. 101% A.mC.mC. A.G. G.fU. G. G. A*fC*fU* A.mC.mU.mU. A. G*fU*fC* A. A.Chl 17392 524 2961mC.mU.mC.mC.mU. 2962 P.mU. G.fU. A.fU. A. 49% A.mU.mU. A.mU. A.fU. A. G.G. A. G* A* A.mC. A.Chl G*G*fU*fU*A. 17393 448 2963 G. A.mU.mC. A.mC.2964 P.mU.fU.fC. A. A. A.fU. 29% A.mU.mU.mU. G. A. G.fU. G. A.fU.fC*fU*G* A.Chl G* A*fU* G. 17394 448 2965 A. G. A.mU.mC. 2966 P.mU.fU.fC. A.A. A.fU. 31% A.mC. G.fU. G. A.fU.fC*fU* G* A.mU.mU.mU. G. A. G* A*fU* G.A.Chl 17395 519 2967 A. 2968 P.mU. A.fU. A. G. G. A. 12%A.mC.mC.mU.mC.mU. G.A. G. G.fU.fU* A* G* mC.mC.mU. A.mU. A* G* A* A.A.Chl 17396 437 2969 G.mU.mU. G. A.mC. 2970 P.mU.fC.fU. G. G. A.fU. 86%A.mU.mC.mC. A. G. G.fU.fC. A. A.fC* A*fC* A.Chl A*fU* A* A. 17397 4062971 mC.mC.mU.mU.mC. 2972 P.mU.fU.fU.fC. G. A. A. 23% mC.mU.mU.mC. G. G.G. A. A. G. G* G* A* A. A. A.Chl A*fU* G* U. 17398 339 2973A.mC.mU.mC.mC. 2974 P.mU.fU. G.fU. 102% A. A. A.mC. A.mC. A. G.fU.fU.fU.G. G. A. A.Chl G.fU* G* G* G*fU*fU* U. 17399 339 2975 mC. 2976 P.mU.fU.G.fU. 55% A.mC.mU.mC.mC. A. G.fU.fU.fU. G. G. A. A. A.mC. A.mC. A. G.fU*G* G* G*fU*fU* A.Chl U. 17400 338 2977 mC. 2978 P.mU. G.fU. G.fU.fU.fU.62% A.mC.mU.mC.mC. A. G. G. A. G.fU. G* G* A. A.mC. A.mC. A.ChlG*fU*fU*fU* C. 17401 468 2979 mC.mC. A.mC.mC. A. 2980 P.mU. G.fU. A. A.61% A.mC.mU.mU. A.mC. G.fU.fU. G. G.fU. G. G* A.Chl A*fC*fU* G*fU* C.17402 468 2981 mU.mC.mC. 2982 P.mU. G.fU. A. A. 179% A.mC.mC. A.G.fU.fU. G. G.fU. G. G* A.mC.mU.mU. A.mC. A*fC*fU* G*fU* C. A.Chl 174031465 2983 A. A.mU. A.mC.mC. 2984 P.mU. A. A. G. A.fC.fU. 30% A. G. G.fU.A.fU.fU*fU*fC* G.mU.mC.mU.mU. A*fU*fC* U. A.Chl 17404 243 2985 G.A.mC.mC. A. 2986 P.mU.fC.fU.fU. A.fU. 32% G.mU. A.mU. A. A. A.fC.fU. G.G.fU.fC* A* G. A.Chl A* A*fU*fC* C. 17405 1472 2987 G.mU.mC.mU.mU.mU.2988 P.mU.fU.fC. A.fU.fU. A. 15% mU. A. A.mU. G. A. A. A. G. A.fC*fU* G*A. A.Chl G*fU* A* U. 17406 2446 2989 A. 2990 P.mU. A. G. A.fC. A.fU.142% A.mU.mU.mU.mC. G.A. A. A.fU.fU* A.mU. A*fC*fU* G* G* U. G.mU.mC.mU.A.Chl 17407 449 2991 A.mU.mC. A.mC. 2992 P.mU. A.fU.fC. A. A. 54%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.Chl A.fU*fC*fU* G* G* A* U. 17408449 2993 G. A.mU.mC. A.mC. 2994 P.mU. A.fU.fC. A. A. 27% A.mU.mU.mU. G.A.fU. G.fU. G. A.mU. A.Chl A.fU*fC*fU* G* G* A* U. 17409 444 2995mU.mC.mC. A. G. 2996 P.mU. A.fU. G.fU. G. 49% A.mU.mC. A.mC. A.fU.fC.fU.G. G. A*fU* A.mU. A.Chl G*fU*fC* A* A. 17410 1093 2997 mU. A.mC.mU. G.2998 P.mU.fC.fU.fC.fC.fU. 32% A.mU. A. G. G.A. G. A.fU.fC. A. G.fU.A.Chl A*fU*fU* A* G*fC* C. 17411 1134 2999 G.mU. G.mC. A. 3000 P.mU.fC.A. A. G.fU. 70% A.mC. A.mC.mU.fU. G.fU.fU. G.mC. A.fC* G. A.Chl A*fU* A*A*fU* C. 17412 244 3001 A.mC.mC. A. G.mU. 3002 P.mU. A.fC.fU.fU. A.fU.63% A.mU. A. A. G.mU. A.fC.fU. G. G.fU*fC* A* A.Chl A* A*fU* C. 174131946 3003 G. A. A. 3004 P.mU.fU.fC. A.fU.fU. A. 19% G.mU.mC.mU. A. G.A.fC.mU.fU.fC*fU* A.mU. G. A. A.Chl A*fC* A* G* U. 17414 638 3005 A. A.G. A. A. G. A. 3006 P.mU. A. 27% A. A. G.mU.mU.A.fC.fU.fU.fU.fC.fU.fU.fC. A.Chl fU.fU* A* G* A* A* G* C. 17415 450 3007mU.mC. A.mC. 3008 P.mU. A. A.fU.fC. A. A. 216% A.mU.mU.mU. G. A.fU.G.fU. G. A.mU.mU. A.Chl A*fU*fC*fU* G* G* A. 17416 450 3009 A.mU.mC.A.mC. 3010 P.mU. A. A.fU.fC. A. A. 32% A.mU.mU.mU. G. A.fU. G.fU. G.A.mU.mU. A.Chl A*fU*fC*fU* G* G* A. 17417 452 3011 A.mC. 3012P.mU.fU.fC. A. A.fU.fC. 99% A.mU.mU.mU. G. A. A. A.fU. G.fU* G* A.mU.mU.G. A. A*fU*fC*fU* G. A.Chl 17418 452 3013 mC. A.mC. 3014 P.mU.fU.fC. A.A.fU.fC. 54% A.mU.mU.mU. G. A. A. A.fU. G.fU* G* A.mU.mU. G. A.A*fU*fC*fU* G. A.Chl 17419 454 3015 A.mU.mU.mU. G. 3016 P.mU.fU.G.fU.fC. A. 86% A.mU.mU. G. A.mC. A.fU.fC. A. A. A.fU* A. A.Chl G*fU* G*A*fU* C. 17420 454 3017 mC. A.mU.mU.mU. 3018 P.mU.fU. G.fU.fC. A. 89% G.A.mU.mU. G. A.fU.fC. A. A. A.fU* A.mC. A. A.Chl G*fU* G* A*fU* C. 174211790 3019 mC. A.mU.mC.mU. 3020 P.mU.fU.fU. A.fU.fU. 55% G.mC. A. A.mU.A. A. G.fC. A. G. A.fU. G* A* A.Chl G* A* G* A* C. 17422 1790 3021mU.mC. 3022 P.mU.fU.fU. A.fU.fU. 62% A.mU.mC.mU. G.mC. G.fC. A. G. A.fU.G* A* A. A.mU. A. A. A.Chl G* A* G* A* C. 21180 448 3023 G. A.mU.mC.A.mC. 3024 P.mU.fU.fC. A.mA. A.fU. 76% A.mU.mU.mU. G. A. G.fU. G.A.mU.mC*mU* A.TEG-Chl G* G* A*mU* G. 21181 448 3025 G. A.mU.mC. A.mC.3026 P.mU.fU.fC. A.mA. A.fU. 37% A.mU.mU.mU. G. A. G.fU. G. A.TEG-ChlA.fU.fC*fU*mG*mG*mA* fU* G. 21182 448 3027 G. A.mU.mC. A.mC. 3028P.mU.fU.fC. A. A. A.fU. 29% A.mU.mU.mU. G.fU. G. A.fU.fC*fU* G*G*mA*mA.TEG-Chl G* A*fU* G. 21183 448 3029 mG*mA*mU.mC. 3030 P.mU.fU.fC.A. A. A.fU. 46% A.mC. G.fU. G. A.fU.fC*fU* G* A.mU.mU.mU. G* A*fU* G.G*mA*mA.TEG-Chl 21184 448 3031 mG*mA*mU.mC.mA. 3032 P.mU.fU.fC. A. A.A.fU. 60% mC.mA.mU.mU.mU. G.fU. G. A.fU.fC*fU* G* mG*mA*mA.TEG- G* A*fU*G. Chl 21185 449 3033 G. A.mU.mC. A.mC. 3034 P.mU. A.fU.fC. A. A. 27%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.fU.fC*fU* G* G* A*fU* G.21186 449 3035 G. A.mU.mC. A.mC. 3036 P.mU. A.fU.fC. A. A. 57%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.mU.mC*mU* G* G* A*mU* G.21187 449 3037 G. A.mU.mC. A.mC. 3038 P.mU. A.fU.fC. A.mA. 54%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.mU.mC*mU* G* G* A*mU* G.21188 449 3039 G. A.mU.mC. A.mC. 3040 P.mU. A.fU.fC. A. A. 66%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.mU.mC*mU*mG*mG* mA*mU*G. 21189 449 3041 G. A.mU.mC. A.mC. 3042 P.mU. A.fU.fC. A.mA. 44%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.mU.mC*mU*mG*mG* mA*mU*G. 21190 449 3043 G. A.mU.mC. A.mC. 3044 P.mU. A.fU.fC. A. A. 52%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.fU.fC*fU*mG*mG*mA* fU*G. 21191 449 3045 G. A.mU.mC. A.mC. 3046 P.mU. A.fU.fC. A.mA. 41%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.fU.fC*fU*mG*mG*mA* fU*G. 21192 449 3047 G. A.mU.mC. A.mC. 3048 P.mU. A.fU.fC. A. A. 98%A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.TEG-Chl A.fU.mC*fU*mG*mG* mA*fU*G. 21193 449 3049 G. A.mU.mC. A.mC. 3050 P.mU. A.fU.fC. A. A. 93%A.mU.mU.mU. G. A.fU. G.fU. G. A*mU*mA.TEG-Chl A.fU*fC*fU* G* G* A* U.21194 449 3051 mG*mA*mU.mC. 3052 P.mU. A.fU.fC. A. A. 119% A.mC. A.fU.G.fU. G. A.mU.mU.mU. G. A.fU*fC*fU* G* G* A* A*mU*mA.TEG-Chl U. 21195449 3053 mG*mA*mU.mC.mA. 3054 P.mU. A.fU.fC. A. A. 292% mC.mA.mU.mU.mU.A.fU. G.fU. G. mG.mA*mU*mA.TEG- A.fU*fC*fU* G* G* A* Chl U. 20620 4493055 G. A.mU.mC. A.mC. 3056 P.mU. A.fU.fC. A. A. 24% A.mU.mU.mU. G.A.fU. G.fU. G. A.mU. A.Chl-TEG A.mU*mC*mU* G* G* A* U. 20621 449 3057 G.A.mU.mC. A.mC. 3058 P.mU. A.fU.fC. A. A. 5% A.mU.mU.mU. G. A.fU. G.fU.G. A.mU. A.Chl-TEG A.mU*fC*mU* G* G* A* U. 20622 449 3059 G. A.mU.mC.A.mC. 3060 P.mU. A. U. C. A. A. A. 25% A.mU.mU.mU. G. U. G. U. G. A.mU.A.Chl-TEG A.mU*mC*mU* G* G* A* U. 20623 449 3061 G. A.mU.mC. A.mC. 3062P.mU. A.fU.fC. A. A. 14% A.mU.mU.mU. G. A.fU. G.fU. G. A.mU. A.Chl-TEGA.mU*mC*mU*mG*mG* mA* U. 20588 448 3063 G. A.mU.mC. A.mC. 3064P.mU.fU.fC. A. A. A.fU. 17% A.mU.mU.mU. G. A. G.fU. G. A.mU.mC*mU*A.Chl-TEG G* G* A*mU* G. 20589 448 3065 G. A.mU.mC. A.mC. 3066P.mU.fU.fC. A. A. A.fU. 40% A.mU.mU.mU. G. A. G.fU. G. A.mU.fC*mU*A.Chl-TEG G* G* A*fU* G. 20590 448 3067 G. A.mU.mC. A.mC. 3068 P.mU. U.C. A. A. A. U. 34% A.mU.mU.mU. G. A. G. U. G. A.mU.mC*mU* A.Chl-TEG G*G* A*mU* G. 20591 448 3069 G. A.mU.mC. A.mC. 3070 P.mU.fU.fC. A. A.A.fU. n/a A.mU.mU.mU. G. A. G.fU. G. A.Chl-TEG A.fU.fC*fU*mG*mG*mA* fU*G.

TABLE 20 Inhibition of gene expression with CTGF sd-rxRNA sequences(Accession Number: NM_001901.2) % remaining mRNA expression Oligo StartSEQ ID SEQ ID (1 uM sd-rxRNA, Number Site NO Sense sequence NO Antisensesequence A549) 13980 1222 3071 A.mC. A. G. G. A. 3072 P.mU. A.fC. 98% A.G. A.mU. G.mU. A.fU.fC.fU.fU.fC.fC.mU. A.Chl G.mU* A* G*mU* A*mC* A.13981 813 3073 G. A. G.mU. G. G. 3074 P.mA. G. G.fC. 82% A. G.mC.G.fC.fU.fC.fC. G.mC.mC.mU.Chl A.mC.mU.mC*mU* G*mU* G* G* U. 13982 7473075 mC. G. A.mC.mU. 3076 P.mU. 116% G. G. A. A. G. G.fU.fC.fU.fU.fC.fC.A. A.mC. A.Chl G.mU.mC. G* G*mU* A* A* G* C. 13983 817 3077 G. G. A.G.mC. 3078 P.mG. A. A.fC. A. G. 97% G.mC.mC.mU. G.fC. G.mU.mU.mC.ChlG.fC.mU.mC.mC* A*mC*mU*mC*mU* G. 13984 1174 3079 G.mC.mC. 3080 P.mC. A.G.fU.fU. 102% A.mU.mU. A.mC. G.fU. A. A.fU. G. A. A.mC.mU. G.mC* A* G*G*mC* G.Chl A* C. 13985 1005 3081 G. A. 3082 P.mA. G.fC.fC. A. G. A.114% G.mC.mU.mU.mU. A. A. G.mC.mU.mC* mC.mU. G. A* A* A*mC*mU* U.G.mC.mU.Chl 13986 814 3083 A. G.mU. G. G. A. 3084 P.mC. A. G. G.fC. 111%G.mC. G.fC.fU.fC.fC. G.mC.mC.mU. A.mC.mU*mC*mU* G.Chl G*mU* G* G. 13987816 3085 mU. G. G. A. 3086 P.mA. A.fC. A. G. G.fC. 102% G.mC.G.fC.fU.mC.mC. G.mC.mC.mU. A*mC*mU*mC*mU* G.mU.mU.Chl G* U. 13988 10013087 G.mU.mU.mU. G. 3088 P.mA. G. A. A. A. 99% A. G.fC.fU.fC. A. A.G.mC.mU.mU.mU. A.mC*mU*mU* G* mC.mU.Chl A*mU* A. 13989 1173 3089 mU.G.mC.mC. 3090 P.mA. G.fU.fU. G.fU. 107% A.mU.mU. A.mC. A. A.fU. G. G.mC.A* A. A.mC.mU.Chl G* G*mC* A*mC* A. 13990 749 3091 A.mC.mU. G. G. 3092P.mC. G.fU. 91% A. A. G. A.mC. G.fU.fC.fU.fU.fC.fC. A. A.mC. G.ChlG.mU*mC* G* G*mU* A* A. 13991 792 3093 A. A.mC.mU. 3094 P.mG. G.A.fC.fC. A. G. 97% G.mC.mC.mU. G. G.fC. A. G.mU.mU* G* G.mU.mC.mC.ChlG*mC*mU*mC* U. 13992 1162 3095 A. G. 3096 P.mC. A. G. G.fC. A.fC. 107%A.mC.mC.mU. A. G. G.mU. G.mU.mC.mU*mU* G.mC.mC.mU. G* A*mU* G* A. G.Chl13993 811 3097 mC. A. G. A. 3098 P.mG.fC. G.fC.fU.fC.fC. 113% G.mU. G.G. A. A.fC.fU.mC.mU. G.mC. G.mC.Chl G*mU* G* G*mU*mC* U. 13994 797 3099mC.mC.mU. G. 3100 P.mG. G.fU.fC.fU. G. n/a G.mU.mC.mC. A. G. A.fC.fC. A.G. G. A.mC.mC.Chl G*mC* A* G*mU*mU* G. 13995 1175 3101 mC.mC. 3102P.mA.fC. A. G.fU.fU. 113% A.mU.mU. A.mC. G.fU. A. A.mU. G. A. A.mC.mU.G*mC* A* G* G*mC* G.mU.Chl A. 13996 1172 3103 mC.mU. 3104 P.mG.fU.fU.G.fU. A. 110% G.mC.mC. A.fU. G. G.mC. A. G* A.mU.mU. A.mC. G*mC* A*mC*A* G. A. A.mC.Chl 13997 1177 3105 A.mU.mU. A.mC. 3106 P.mG. G. A.fC. A.105% A. A.mC.mU. G.fU.fU. G.fU. A. G.mU.mC.mC.Chl A.mU* G* G*mC* A* G*G. 13998 1176 3107 mC. A.mU.mU. 3108 P.mG. A.fC. A. G.fU.fU. 89% A.mC.A. G.fU. A. A.mU. G* A.mC.mU. G*mC* A* G* G* C. G.mU.mC.Chl 13999 8123109 A. G. A. G.mU. G. 3110 P.mG. G.fC. 99% G. A. G.mC. G.fC.fU.fC.fC.G.mC.mC.Chl A.fC.mU.mC.mU* G*mU* G* G*mU* C. 14000 745 3111 A.mC.mC. G.3112 P.mU.fC.fU.fU.fC.fC. A. n/a A.mC.mU. G. G. A. G.fU.fC. G. G.mU* A*A. G. A.Chl A* G*mC*mC* G. 14001 1230 3113 A.mU. G.mU. 3114 P.mU. 106%A.mC. G. G. A. G. G.fU.fC.fU.fC.fC. G.fU. A.mC. A.Chl A.mC.A.mU*mC*mU*mU* mC*mC* U. 14002 920 3115 G.mC.mC.mU.mU. 3116 P.mA.G.fC.fU.fU.fC. 93% G.mC. G. A. A. G.fC. A. A. G. G.mC.mU.Chl G.mC*mC*mU*G* A*mC* C. 14003 679 3117 G.mC.mU. G.mC. 3118 P.mC. 102% G. A. G. G. A.A.fC.fU.fC.fC.fU.fC. G.mU. G.Chl G.fC. A. G.mC* A*mU*mU*mU*mC* C. 14004992 3119 G.mC.mC.mU. 3120 P.mA. A. A.fC.fU.fU. G. 100% A.mU.mC. A. A.A.fU. A. G. G.mU.mU.mU.Chl G.mC*mU*mU* G* G* A* G. 14005 1045 3121 A.3122 P.mA.fC.fU.fC.fC. A.fC. 104% A.mU.mU.mC.mU. A. G. A. G.mU. G. G. A.A.mU.mU*mU* A* G.mU.Chl G*mC*mU* C. 14006 1231 3123 mU. G.mU. A.mC. 3124P.mA.fU. 87% G. G. A. G. A.mC. G.fU.fC.fU.fC.fC. G.fU. A.mU.Chl A.mC.A*mU*mC*mU*mU* mC* C. 14007 991 3125 A. G.mC.mC.mU. 3126 P.mA.A.fC.fU.fU. G. 101% A.mU.mC. A. A. A.fU. A. G. G.mU.mU.Chl G.mC.mU*mU*G* G* A* G* A. 14008 998 3127 mC. A. A. 3128 P.mA. A. G.fC.fU.fC. A. 98%G.mU.mU.mU. G. A. A.fC.mU.mU. G* A. A*mU* A* G* G* C. G.mC.mU.mU.Chl14009 1049 3129 mC.mU. G.mU. G. 3130 P.mA.fC. A.fU. 98% G. A. G.mU.A.mU. A.fC.fU.fC.fC. A.mC. A. G.mU.Chl G* A* A*mU*mU*mU* A. 14010 10443131 A. A. 3132 P.mC.fU.fC.fC. A.fC. A. 93% A.mU.mU.mC.mU. G. A.A.mU.mU.mU* G.mU. G. G. A. A* G*mC*mU*mC* G. G.Chl 14011 1327 3133mU.mU.mU.mC. 3134 P.mU. G.fU. G.fC.fU. 95% A. G.mU. A. G.mC. A.fC.fU. G.A. A. A.mC. A.Chl A*mU*mC* A*mU*mU* U. 14012 1196 3135 mC. A. A.mU. G.3136 P.mA. A. A. G. A.fU. 101% A.mC. G.fU.fC. A.mU.mU. A.mU.mC.mU.mU.G*mU*mC*mU*mC* mU.Chl mC* G. 14013 562 3137 A. G.mU. 3138 P.mG.fU. G.fC.A.fC.fU. 66% A.mC.mC. A. G. G.fU. G.mU. G.mC. A.mC.mU*mU* A.mC.Chl G*mC*A* G* C. 14014 752 3139 G. G. A. A. G. 3140 P.mA. A. A.fC. G.fU. 95%A.mC. A.mC. G.fU.fC.fU.mU.mC.mC* G.mU.mU.mU.Chl A* G*mU*mC* G* G. 14015994 3141 mC.mU. 3142 P.mU.fC. A. A. 85% A.mU.mC. A. A. A.fC.fU.fU. G.A.mU. G.mU.mU.mU. G. A. G* A.Chl G*mC*mU*mU* G* G. 14016 1040 3143 A.G.mC.mU. A. 3144 P.mA.fC. A. G. A. 61% A. A.fU.fU.fU. A. A.mU.mU.mC.mU.G.mC.mU*mC* G* G.mU.Chl G*mU* A* U. 14017 1984 3145 A. G. G.mU. A. G.3146 P.mU.fU. A.fC. 32% A. A.mU. G.mU. A. A.fU.fU.fC.fU. A.ChlA.mC.mC.mU* A*mU* G* G*mU* G. 14018 2195 3147 A. G.mC.mU. G. 3148 P.mA.A. A.fC.fU. G. 86% A.mU.mC. A. A.fU.fC. A. G.mC.mU* G.mU.mU.mU.Chl A*mU*A*mU* A* G. 14019 2043 3149 mU.mU.mC.mU. 3150 P.mU. A.fU.fC.fU. G. A.81% G.mC.mU.mC. A. G.fC. A. G. A. G. A.mU. A.Chl A*mU*mU*mU*mC* mC* A.14020 1892 3151 mU.mU. 3152 P.mU.fU. A. 84% A.mU.mC.mU. A. A.fC.fU.fU.A. G. A. G.mU.mU. A. A.mU. A. A*mC*mU* A.Chl G*mU* A* C. 14021 1567 3153mU. A.mU. A.mC. 3154 P.mU. A.fU.fU. 72% G. A. G.mU. A. A.fC.fU.fC. G.fU.A.mU. A.Chl A.mU. A* A* G* A*mU* G* C. 14022 1780 3155 G. A.mC.mU. G.3156 P.mA. A. G.fC.fU. 65% G. A.mC. A. G.fU.fC.fC. A. G.mC.mU.mU.ChlG.mU.mC*mU* A* A*mU*mC* G. 14023 2162 3157 A.mU. G. 3158 P.mU. A. A.fU.A. A. A. 80% G.mC.mC.mU.mU. G. G.fC.mC. mU. A.mU.mU. A.mU*mU*mU* A.ChlG*mU*mU* C. 14024 1034 3159 A.mU. A.mC.mC. 3160 P.mU.fU.fU. A. 91% G. A.G.mC.mU. A. G.fC.fU.fC. G. G.mU. A. A.Chl A.mU* G*mU*mC*mU*mU* C. 140252264 3161 mU.mU. 3162 P.mA.fC. 58% G.mU.mU. G. A. A.fC.fU.fC.fU.fC. A.G. A. G.mU. A.mC. A. A* A*mU* G.mU.Chl A* A* A* C. 14026 1032 3163 A.mC.A.mU. 3164 P.mU. A. G.fC.fU.fC. G. 106% A.mC.mC. G. A. G.fU. A.mU.G.mC.mU. A.Chl G.mU*mC*mU*mU* mC* A* U. 14027 1535 3165 A. G.mC. A. G.A. 3166 P.mU. A. 67% A. A. G. G.mU.mU. A.fC.fC.fU.fU.fU.fC.fU. A.ChlG.mC.mU* G* G*mU* A*mC* C. 14028 1694 3167 A. G.mU.mU. 3168 P.mU.fU. A.A. G. G. A. 94% G.mU.mU.mC.mC. A.fC. A. mU.mU. A. A.Chl A.mC.mU*mU* G*A*mC*mU* C. 14029 1588 3169 A.mU.mU.mU. G. 3170 P.mU.fU. A.fC. 97% A. A.G.mU. G.mU. A.fC.fU.fU.fC. A. A. A. A.Chl A.mU* A* G*mC* A* G* G. 14030928 3171 A. A. G.mC.mU. 3172 P.mU.fC.fC. A. G. 100% G. A.mC.mC.mU.G.fU.fC. A. G. G. A.Chl G.mC.mU.mU*mC* G*mC* A* A* G. 14031 1133 3173 G.G.mU.mC. 3174 P.mC.fU.fU.fC.fU.fU.fC. 82% A.mU. G. A. A. G. A.fU. G. A.A. G.Chl A.mC.mC*mU*mC* G*mC*mC* G. 14032 912 3175 A.mU. G. 3176 P.mA.A. G. G.fC.fC.fU. 84% G.mU.mC. A. G. G. A.fC.mC. A.mU* G.mC.mC.mU.mU.G*mC* A*mC* A* G. Chl 14033 753 3177 G. A. A. G. A.mC. 3178 P.mC. A. A.A.fC. G.fU. 86% A.mC. G.fU.fC.mU.mU.mC*mC* G.mU.mU.mU. A* G*mU*mC* G.G.Chl 14034 918 3179 A. G. 3180 P.mC.fU.fU.fC. G.fC. A. 88%G.mC.mC.mU.mU. A. G. G.mC.mC.mU* G.mC. G. A. A. G* A*mC*mC* A* U. G.Chl14035 744 3181 mU. A.mC.mC. G. 3182 P.mC.fU.fU.fC.fC. A. 95% A.mC.mU. G.G. A. G.fU.fC. G. G.mU. A* A. G.Chl A* G*mC*mC* G* C. 14036 466 3183A.mC.mC. G.mC. 3184 P.mC.fC. G. 73% A. A. G. A.mU.mC. A.fU.fC.fU.fU.G.fC. G. G. G.Chl G.mU*mU* G* G*mC*mC* G. 14037 917 3185 mC. A. G. 3186P.mU.fU.fC. G.fC. A. A. 86% G.mC.mC.mU.mU. G. G.fC.mC.mU. G* G.mC. G. A.A.Chl A*mC*mC* A*mU* G. 14038 1038 3187 mC. G. A. 3188 P.mA. G. A. 84%G.mC.mU. A. A. A.fU.fU.fU. A. A.mU.mU.mC.mU. G.fC.mU.mC. G* Chl G*mU*A*mU* G* U. 14039 1048 3189 mU.mC.mU. 3190 P.mC. A.fU. 87% G.mU. G. G.A. A.fC.fU.fC.fC. A.fC. A. G.mU. A.mU. G. A* G.Chl A*mU*mU*mU* A* G.14040 1235 3191 mC. G. G. A. G. 3192 P.mU. G.fC.fC. A.fU. 100% A.mC.A.mU. G. G.fU.fC.fU.mC.mC. G.mC. A.Chl G*mU* A*mC* A*mU* C. 14041 8683193 A.mU. G. A.mC. 3194 P.mG. A. G. G.fC. 104% A. A.mC. G.fU.fU.G.fU.mC. G.mC.mC.mU.mC. A.mU*mU* G* Chl G*mU* A* A. 14042 1131 3195 G.A. G. 3196 P.mU.fC.fU.fU.fC. 85% G.mU.mC. A.mU. A.fU. G. G. A. A. G.A.Chl A.fC.mC.mU.mC* G*mC*mC* G*mU* C. 14043 1043 3197 mU. A. A. 3198P.mU.fC.fC. A.fC. A. G. 74% A.mU.mU.mC.mU. A. A.fU.mU.mU. A* G.mU. G. G.A.Chl G*mC*mU*mC* G* G. 14044 751 3199 mU. G. G. A. A. G. 3200 P.mA.A.fC. G.fU. 84% A.mC. A.mC. G.fU.fC.fU.fU.mC.mC. G.mU.mU.Chl A* G*mU*mC*G* G* U. 14045 1227 3201 A. A. G. A.mU. 3202 P.mC.fU.fC.fC. G.fU. 99%G.mU. A.mC. G. G. A.fC. A. G.Chl A.fU.mC.mU.mU*mC* mC*mU* G*mU* A. 14046867 3203 A. A.mU. G. 3204 P.mA. G. G.fC. G.fU.fU. 94% A.mC. A. A.mC.G.fU.fC. A.mU.mU* G* G.mC.mC.mU.Chl G*mU* A* A* C. 14047 1128 3205 G.G.mC. G. A. G. 3206 P.mU.fC. A.fU. G. 89% G.mU.mC. A.mU. A.fC.fC.fU.fC.G. A.Chl G.mC.mC* G*mU*mC* A* G* G. 14048 756 3207 G. A.mC. A.mC. 3208P.mG. G.fC.fC. A. A. 93% G.mU.mU.mU. G. A.fC. G.fU. G.mC.mC.ChlG.mU.mC*mU*mU*mC* mC* A* G. 14049 1234 3209 A.mC. G. G. A. G. 3210P.mG.fC.fC. A.fU. 100% A.mC. A.mU. G. G.fU.fC.fU.fC.mC. G.mC.Chl G.mU*A*mC* A*mU*mC* U. 14050 916 3211 mU.mC. A. G. 3212 P.mU.fC. G.fC. A. A.G. 96% G.mC.mC.mU.mU. G.fC.fC.mU. G. G.mC. G. A.Chl A*mC*mC* A*mU* G* C.14051 925 3213 G.mC. G. A. A. 3214 P.mA. G. G.fU.fC. A. 80% G.mC.mU. G.G.fC.fU.fU.mC. G.mC* A.mC.mC.mU.Chl A* A* G* G*mC* C. 14052 1225 3215 G.G. A. A. G. 3216 P.mC.fC. G.fU. A.fC. 96% A.mU. G.mU.A.fU.fC.fU.mU.mC.mC* A.mC. G. G.Chl mU* G*mU* A* G* U. 14053 445 3217G.mU. G. 3218 P.mG. A. G.fC.fC. G. A. 101% A.mC.mU.mU.mC. A. G.fU.mC.A.mC* A* G. G* A* A* G* A. G.mC.mU.mC.Chl 14054 446 3219 mU. G. 3220P.mG. G. A. G.fC.fC. G. 93% A.mC.mU.mU.mC. A. A. G.mU.mC. G. A*mC* A* G*A* A* G.mC.mU.mC.mC. G. Chl 14055 913 3221 mU. G. G.mU.mC. 3222 P.mC. A.A. G. 67% A. G. G.fC.fC.fU. G. G.mC.mC.mU.mU. A.mC.mC. A*mU* G.Chl G*mC*A*mC* A. 14056 997 3223 mU.mC. A. A. 3224 P.mA. G.fC.fU.fC. A. A. 92%G.mU.mU.mU. G. A.fC.fU.mU. G. A*mU* A. G.mC.mU.Chl A* G* G*mC* U. 14057277 3225 G.mC.mC. A. G. 3226 P.mC.fU. G.fC. A. 84% A. A.mC.mU.G.fU.fU.fC.fU. G. G.mC. A. G.Chl G.mC*mC* G* A*mC* G* G. 14058 1052 3227mU. G. G. A. 3228 P.mG. G.fU. A.fC. A.fU. n/a G.mU. A.mU. A.fC.fU.mC.mC.G.mU. A*mC* A* G* A* A* A.mC.mC.Chl U. 14059 887 3229 G.mC.mU. A. G.3230 P.mC.fU. 80% A. G. A. A. G.mC. G.fC.fU.fU.fC.fU.fC.fU. A. G.Chl A.G.mC*mC*mU* G*mC* A* G. 14060 914 3231 G. G.mU.mC. A. 3232 P.mG.fC. A.A. G. 112% G. G.fC.fC.fU. G. G.mC.mC.mU.mU. A.mC.mC* A*mU* G.mC.ChlG*mC* A* C. 14061 1039 3233 G. A. G.mC.mU. 3234 P.mC. A. G. A. 104% A.A. A.fU.fU.fU. A. A.mU.mU.mC.mU. G.mC.mU.mC* G* G.Chl G*mU* A*mU* G.14062 754 3235 A. A. G. A.mC. 3236 P.mC.fC. A. A. A.fC. 109% A.mC. G.fU.G.mU.mU.mU. G. G.fU.mC.mU.mU*mC* G.Chl mC* A* G*mU* C. 14063 1130 3237mC. G. A. G. 3238 P.mC.fU.fU.fC. A.fU. G. 103% G.mU.mC. A.mU.A.fC.fC.mU.mC. G. A. A. G.Chl G*mC*mC* G*mU*mC* A. 14064 919 3239 G.3240 P.mG.fC.fU.fU.fC. 109% G.mC.mC.mU.mU. G.fC. A. A. G. G.mC. G. A. A.G.mC.mC*mU* G* G.mC.Chl A*mC*mC* A. 14065 922 3241 mC.mU.mU. 3242P.mU.fC. A. 106% G.mC. G. A. A. G.fC.fU.fU.fC. G.fC. A. G.mC.mU. G. A.G* G*mC*mC*mU* A.Chl G* A. 14066 746 3243 mC.mC. G. 3244P.mG.fU.fC.fU.fU.fC.fC. 106% A.mC.mU. G. G. A. A. G.fU.mC. G. A. G.A.mC.Chl G*mU* A* A* G*mC* C. 14067 993 3245 mC.mC.mU. 3246 P.mC. A. A.A.fC.fU.fU. 67% A.mU.mC. A. A. G. A.fU. A. G. G.mU.mU.mU. G*mC*mU*mU* G*G.Chl G* A. 14068 825 3247 mU. 3248 P.mA. G. 93% G.mU.mU.mC.mC.G.fU.fC.fU.fU. G. G. A. A. A. G. A.mC. A* G* G*mC* A.mC.mC.mU.Chl G*mC*U. 14069 926 3249 mC. G. A. A. 3250 P.mC. A. G. G.fU.fC. A. 95% G.mC.mU.G. G.fC.fU.mU.mC. A.mC.mC.mU. G*mC* A* A* G* G* G.Chl C. 14070 923 3251mU.mU. G.mC. G. 3252 P.mG.fU.fC. A. 95% A. A. G.mC.mU. G. G.fC.fU.fU.fC.G.mC. A. A.mC.Chl A* G* G*mC*mC*mU* G. 14071 866 3253 mC. A. A.mU. G.3254 P.mG. G.fC. G.fU.fU. 132% A.mC. A. A.mC. G.fU.fC. A.mU.mU. G*G.mC.mC.Chl G*mU* A* A*mC* C. 14072 563 3255 G.mU. A.mC.mC. 3256 P.mC.G.fU. G.fC. n/a A. G.mU. G.mC. A.fC.fU. G. G.mU. A.mC. G.Chl A.mC*mU*mU*G*mC* A* G. 14073 823 3257 mC.mC.mU. 3258 P.mG.fU.fC.fU.fU. G. 98%G.mU.mU.mC.mC. G. A. A.fC. A. G. A. A. G. A.mC.Chl G*mC* G*mC*mU*mC* C.14074 1233 3259 mU. A.mC. G. G. 3260 P.mC.fC. A.fU. 109% A. G. A.mC.A.mU. G.fU.fC.fU.fC.fC. G. G.Chl G.mU. A*mC* A*mU*mC*mU* U. 14075 9243261 mU. G.mC. G. A. 3262 P.mG. G.fU.fC. A. 95% A. G.mC.mU. G.G.fC.fU.fU.fC. G.mC. A.mC.mC.Chl A* A* G* G*mC*mC* U. 14076 921 3263mC.mC.mU.mU. 3264 P.mC. A. G.fC.fU.fU.fC. 116% G.mC. G. A. A. G.fC. A.A. G. G.mC.mU. G.Chl G*mC*mC*mU* G* A* C. 14077 443 3265 mC.mU. G.mU. G.3266 P.mG.fC.fC. G. A. A. 110% A.mC.mU.mU.mC. G.fU.fC. A.mC. A. G* G.G.mC.Chl A* A* G* A* G* G. 14078 1041 3267 G.mC.mU. A. A. 3268 P.mC.A.fC. A. G. A. 99% A.mU.mU.mC.mU. A.fU.fU.fU. A. G.mU. G.Chl G.mC*mU*mC*G* G*mU* A. 14079 1042 3269 mC.mU. A. A. 3270 P.mC.fC. A.fC. A. G. A.109% A.mU.mU.mC.mU. A.fU.fU.mU. A. G.mU. G. G.Chl G*mC*mU*mC* G* G* U.14080 755 3271 A. G. A.mC. A.mC. 3272 P.mG.fC.fC. A. A. A.fC. 121%G.mU.mU.mU. G. G.fU. G.mC.Chl G.mU.mC.mU*mU*mC* mC* A* G* U. 14081 4673273 mC.mC. G.mC. A. 3274 P.mG.fC. C.fG. A. 132% A. G. A.mU.mC. G.U.fC.fU.fU.fG. C.mG. G.mC.Chl G*mU*mU* G* G*mC* C. 14082 995 3275 mU.A.mU.mC. A. 3276 P.mC.fU.fC. A. A. 105% A. G.mU.mU.mU. A.fC.fU.fU. G.A.mU. G. A. G.Chl A* G* G*mC*mU*mU* G. 14083 927 3277 G. A. A. 3278P.mC.fC. A. G. G.fU.fC. 114% G.mC.mU. G. A. G.fC.mU.mU.mC* A.mC.mC.mU.G. G*mC* A* A* G* G. G.Chl 17356 1267 3279 A.mC. A.mU.mU. 3280 P.mU.A.fU. G. A. 120% A. A.mC.mU.mC. G.mU.fU. A. A.fU. A.mU. A.ChlG.fU*fC*fU*fC*fU*fC* A. 17357 1267 3281 G. A.mC. 3282 P.mU. A.fU. G. A.56% A.mU.mU. A. G.mU.fU. A. A.fU. A.mC.mU.mC. G.fU*fC*fU*fC*fU*fC* A.mU.A.Chl A. 17358 1442 3283 mU. G. A. A. G. A. 3284 P.mU.fU. A. A.fC. 34%A.mU. G.mU.mU. A.fU.fU.fC.fU.fU.fC. A* A. A.Chl A* A*fC*fC* A* G. 173591442 3285 mU.mU. G. A. A. 3286 P.mU.fU. A. A.fC. 31% G. A. A.mU.A.fU.fU.fC.fU.fU.fC. A* G.mU.mU. A. A* A*fC*fC* A* G. A.Chl 17360 15573287 G. A.mU. A. 3288 P.mU.fU. A. A. G. A.fU. 59% G.mC. G.fC.fU.A.fU.fC*fU* A.mU.mC.mU.mU. G* A*fU* G* A. A. A.Chl 17361 1557 3289 A. G.A.mU. A. 3290 P.mU.fU. A. A. G. A.fU. 47% G.mC. G.fC.fU. A.fU.fC*fU*A.mU.mC.mU.mU. G* A*fU* G* A. A. A.Chl 17362 1591 3291 mU. G. A. A. 3292P.mU. A. A.fU.fU. A.fC. 120% G.mU. G.mU. A. A.fC.fU.fU.fC. A* A*A.mU.mU. A.Chl A*fU* A* G* C. 17363 1599 3293 A. A.mU.mU. G. 3294P.mU.fU.fC.fC.fU.fU.fC. 71% A. G. A. A. G. G. A. fU.fC. A. A.fU.fU*A.Chl A*fC* A*fC*fU* U. 17364 1601 3295 mU.mU. G. A. G. 3296P.mU.fU.fU.fU.fC.fC.fU. 62% A. A. G. G. A. A. A. fU.fC.fU.fC. A. A.ChlA*fU*fU* A*fC* A* C. 17365 1732 3297 mC. 3298 P.mU.fC. G. A. A.fU.fC.99% A.mU.mU.mC.mU. A. G. A. A.fU. G. A.mU.mU.mC. G*fU*fC* A* G* A* G. G.A.Chl 17366 1734 3299 mU.mU.mC.mU. 3300 P.mU.fU.fU.fC. G. A. 97% G.A.mU.mU.mC. A.fU.fC. A. G. A. A*fU* G. A. A. A.Chl G*fU*fC* A* G. 173671770 3301 mC.mU. 3302 P.mU.fU.fC.fU. A. 45% G.mU.mC. G. A.fU.fC. G.A.fC. A. G* A.mU.mU. A. G. A. G* A*fU*fU*fC* C. A.Chl 17368 1805 3303mU.mU.mU. 3304 P.mU. G.fU.fU. A.fC. A. 71% G.mC.mC.mU. G. G.fC. A. A.G.mU. A. A.mC. A*fU*fU*fC* A*fC* U. A.Chl 17369 1805 3305 A.mU.mU.mU.3306 P.mU. G.fU.fU. A.fC. A. 67% G.mC.mC.mU. G. G.fC. A. A. G.mU. A.A.mC. A*fU*fU*fC* A*fC* U. A.Chl 17370 1815 3307 A.mC. A. A. 3308 P.mU.A. A.fU.fC.fU. G. 65% G.mC.mC. A. G. G.fC.fU.fU. G.fU*fU* A.mU.mU. A.ChlA*fC* A* G* G. 17371 1815 3309 A. A.mC. A. A. 3310 P.mU. A. A.fU.fC.fU.G. 35% G.mC.mC. A. G. G.fC.fU.fU. G.fU*fU* A.mU.mU. A.Chl A*fC* A* G* G.17372 2256 3311 mC. A. 3312 P.mU. A.fC. A. A. A.fU. 113% G.mU.mU.mU. A.A. A.fC.fU. A.mU.mU.mU. G*fU*fC*fC* G* A* A. G.mU. A.Chl 17373 2265 3313mU. G.mU.mU. G. 3314 P.mU. A.fC. 35% A. G. A. G.mU. A.fC.fU.fC.fU.fC. A.G.mU. A.Chl A.fC. A* A* A*fU* A* A* A. 17374 2265 3315 mU.mU. 3316 P.mU.A.fC. 31% G.mU.mU. G. A. A.fC.fU.fC.fU.fC. A. G. A. G.mU. A.fC. A* A*A*fU* A* G.mU. A.Chl A* A. 17375 2295 3317 mU. G.mC. 3318 P.mU.fU. A. G.A. A. A. 34% A.mC.mC.mU.mU. G. G.fU. G.fC. A* A* mU.mC.mU. A. A*fC*A*fU* G. A.Chl 17376 2295 3319 mU.mU. G.mC. 3320 P.mU.fU. A. G. A. A. A.28% A.mC.mC.mU.mU. G. G.fU. G.fC. A* A* mU.mC.mU. A. A*fC* A*fU* G.A.Chl 17377 1003 3321 mU.mU. G. A. 3322 P.mU.fC. A. G. A. A. A. 67%G.mC.mU.mU.mU. G.fC.fU.fC. A. A* mC.mU. G. A.Chl A*fC*fU*fU* G* A. 173782268 3323 mU. G. A. G. A. 3324 P.mU. G.fU.fC. A.fC. 42% G.mU. G.mU. G.A.fC.fU.fC.fU.fC. A* A.mC. A.Chl A*fC* A* A* A* U. 17379 2272 3325 A.G.mU. G.mU. 3326 P.mU.fU.fU.fU. G. 35% G. A.mC.mC. A. A. G.fU.fC. A.fC.A. A.Chl A.fC.fU*fC*fU*fC* A* A* C. 17380 2272 3327 G. A. G.mU. 3328P.mU.fU.fU.fU. G. 29% G.mU. G. G.fU.fC. A.fC. A.mC.mC. A. A. A.A.fC.fU*fC*fU*fC* A* A.Chl A* C. 17381 2273 3329 G.mU. G.mU. G. 3330P.mU.fU.fU.fU.fU. G. 42% A.mC.mC. A. A. A. G.fU.fC. A.fC. A. A.ChlA.fC*fU*fC*fU*fC* A* A. 17382 2274 3331 mU. G.mU. G. 3332P.mU.fC.fU.fU.fU.fU. 42% A.mC.mC. A. A. A. G. G.fU.fC. A.fC. A. G. A.ChlA*fC*fU*fC*fU*fC* A. 17383 2274 3333 G.mU. G.mU. G. 3334P.mU.fC.fU.fU.fU.fU. 37% A.mC.mC. A. A. A. G. G.fU.fC. A.fC. A. G. A.ChlA*fC*fU*fC*fU*fC* A. 17384 2275 3335 G.mU. G. 3336 P.mU. 24% A.mC.mC. A.A. A. A.fC.fU.fU.fU.fU. G. A. G.mU. A.Chl G.fU.fC. A.fC* A*fC*fU*fC*fU*C. 17385 2277 3337 G. A.mC.mC. A. A. 3338 P.mU.fU. A. 27% A. A. G.mU.mU.A. A.fC.fU.fU.fU.fU. G. A.Chl G.fU.fC* A*fC* A*fC*fU* C. 17386 2296 3339G.mC. 3340 P.mU.fC.fU. A. G. A. A. 23% A.mC.mC.mU.mU. A. G. G.fU. G.fC*A* mU.mC.mU. A. G. A* A*fC* A* U. A.Chl 17387 2299 3341 mC.mC.mU.mU.mU.3342 P.mU.fC. A. A.fC.fU. A. 46% mC.mU. A. G. A. A. A. G. G*fU* G.mU.mU.G. G*fC* A* A* A. A.Chl 21138 2296 3343 G.mC. 3344 P.mU.fC.fU. A. G. 42%A.mC.mC.mU.mU. A.mA. A. G. G.fU. mU.mC.mU. A. G. G.mC* A* A* A*mC*A.TEG-Chl A* U. 21139 2296 3345 G.mC. 3346 P.mU.fC.fU. A. G.mA. 32%A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU. A. G. A* A* A*mC* A* U.A.TEG-Chl 21140 2296 3347 G.mC. 3348 P.mU.fC.fU. A. G. A. A. 41%A.mC.mC.mU.mU. A. G. G.fU. G.mC* mU.mC.mU. A. G. A*mA* A*mC* A* U.A.TEG-Chl 21141 2296 3349 G.mC. 3350 P.mU.fC.fU. A. G. 51%A.mC.mC.mU.mU. A.mA. A. G. G.fU. mU.mC.mU. A. G. G.mC* A*mA* A*mC*A.TEG-Chl A* U. 21142 2296 3351 G.mC. 3352 P.mU.fC.fU. A. G.mA. 25%A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU. A. G. A*mA* A*mC* A* U.A.TEG-Chl 21143 2296 3353 G.mC. 3354 P.mU.fC.fU. A. G. A. A. 61%A.mC.mC.mU.mU. A. G. G.fU. mU.mC.mU. A. G. G.fC*mA*mA*mA*fC* A.TEG-ChlmA* U. 21144 2296 3355 G.mC. 3356 P.mU.fC.fU. A. G. 49% A.mC.mC.mU.mU.A.mA. A. G. G.fU. mU.mC.mU. A. G. G.fC*mA*mA*mA*fC* A.TEG-Chl mA* U.21145 2296 3357 G.mC. 3358 P.mU.fC.fU. A. G.mA. 46% A.mC.mC.mU.mU. A.mA.G. G.fU. mU.mC.mU. A. G. G.fC*mA*mA*mA*fC* A.TEG-Chl mA* U. 21146 22963359 G.mC. 3360 P.mU.fC.fU. A. G. A. A. 37% A.mC.mC.mU.mU. A. G. G.fU.G.fC* A* mU.mC.mU. A* A*fC* A* U. A*mG*mA.TEG- Chl 21147 2296 3361mG*mC* 3362 P.mU.fC.fU. A. G. A. A. 43% A.mC.mC.mU.mU. A. G. G.fU. G.fC*A* mU.mC.mU. A* A*fC* A* U. A*mG*mA.TEG- Chl 21148 2296 3363mG*mC*mA.mC. 3364 P.mU.fC.fU. A. G. A. A. 29% mC.mU.mU.mU.mC. A. G.G.fU. G.fC* A* mU.mA*mG*mA. A* A*fC* A* U. TEG-Chl 21149 2275 3365 G.mU.G. 3366 P.mU. 138% A.mC.mC. A. A. A. A.fC.fU.fU.fU.fU. G. A. G.fU.fC.A.fC* G*mU*mA.TEG- A*fC*fU*fC*fU* C. Chl 21150 2275 3367 mG*mU* G. 3368P.mU. 116% A.mC.mC. A. A.fC.fU.fU.fU.fU. G. A.mA. A. G.fU.fC. A.fC*G*mU*mA.TEG- A*fC*fU*fC*fU* C. Chl 21151 2275 3369 mG*mU*mG.mA. 3370P.mU. 105% mC.mC.mA.mA.mA. A.fC.fU.fU.fU.fU. G. mA.mG*mU*mA. G.fU.fC.A.fC* TEG-Chl A*fC*fU*fC*fU* C. 21152 2295 3371 mU.mU. G.mC. 3372P.mU.fU. A. G. A.mA. 46% A.mC.mC.mU.mU. A. G. G.fU. G.fC. A. A*mU.mC.mU. A. A*fC* A*fA* G* G. A.TEG-Chl 21153 2295 3373 mU.mU. G.mC.3374 P.mU.fU. A. G.mA. 28% A.mC.mC.mU.mU. A.mA. G. G.fU. G.fC. A.mU.mC.mU. A. A* A*fC* A*fA* G* G. A.TEG-Chl 21154 2295 3375 mU.mU. G.mC.3376 P.mU.fU.mA. G.mA. 28% A.mC.mC.mU.mU. A.mA. G.mG.fU. G.fC. mU.mC.mU.A. A. A* A*fC* A*fA* G* A.TEG-Chl G. 21155 2295 3377 mU.mU. G.mC. 3378P.mU.fU. A. G. A.mA. 60% A.mC.mC.mU.mU. A. G. G.fU. G.mC. A. mU.mC.mU.A. A* A*mC* A*mA* G* A.TEG-Chl G. 21156 2295 3379 mU.mU. G.mC. 3380P.mU.fU. A. G. A.mA. 54% A.mC.mC.mU.mU. A. G. G.fU. G.fC. mU.mC.mU. A.A.mA*mA*fC*mA*fA* A.TEG-Chl mG* G. 21157 2295 3381 mU.mU. G.mC. 3382P.mU.fU. A. G. A.mA. 40% A.mC.mC.mU.mU. A. G. G.fU. mU.mC.mU. A.G.fC.mA.mA*mA*fC* A.TEG-Chl mA*fA*mG* G. 21158 2295 3383 mU.mU. G.mC.3384 P.mU.fU. A. G. A.mA. n/a A.mC.mC.mU.mU. A. G. G.fU. G.fC. mU.mC.mU.A. A.mA*mA*fC*mA*mA* A.TEG-Chl mG* G. 21159 2295 3385 mU.mU. G.mC. 3386P.mU.fU. A. G. A.mA. 41% A.mC.mC.mU.mU. A. G. G.fU. G.fC. mU.mC.mU. A.A.mA*mA*mC*mA*mA* A.TEG-Chl mG* G. 21160 2295 3387 mU.mU. G.mC. 3388P.mU.fU. A. G. A.mA. 65% A.mC.mC.mU.mU. A. G. G.fU. G.fC.mA. mU.mC.mU.A. A*mA*mC*mA*mA* A.Chl-TEG mG*mG. 21161 2295 3389 mU.mU. G.mC. 3390P.mU.fU. A. G. A.mA. 43% A.mC.mC.mU.mU. A. G. G.fU. G.fC. A. A*mU.mC.mU. A. A*fC* A*mA*mG* G. A.TEG-Chl 21162 2295 3391 mU.mU. G.mC.3392 P.mU.fU. A. G. A.mA. 41% A.mC.mC.mU.mU. A. G. G.fU. G.fC.mA.mU.mC.mU. A. A*mA*fC* A.TEG-Chl A*mA*mG* G. 21163 2295 3393 mU.mU. G.mC.3394 P.mU.fU. A. G. A. A. A. 32% A.mC.mC.mU.mU. G. G.fU. G.fC. A. A*mU.mC.mU. A* A*fC* A* A* G* G. A*TEG-Chl 21164 2295 3395 mU.mU. G.mC.3396 P.mU.fU. A. G. A. A. A. 39% A.mC.mC.mU.mU. G. G.fU. G.fC. A. A*mU.mC.mU.mA* A*fC* A* A* G* G. mA*TEG-Chl 21165 2295 3397 mU*mU* G.mC.3398 P.mU.fU. A. G. A. A. A. 28% A.mC.mC.mU.mU. G. G.fU. G.fC. A. A*mU.mC.mU.mA* A*fC* A* A* G* G. mA*TEG-Chl 21166 2295 3399mU.mU.mG.mC.mA. 3400 P.mU.fU. A. G. A. A. A. 27% mC.mC.mU.mU. G. G.fU.G.fC. A. A* mU.mC.mU.mA* A*fC* A* A* G* G. mA*TEG-Chl 21167 2299 3401mC.mC.mU.mU.mU. 3402 P.mU.fC. A. A.fC.fU. A. 49% mC.mU. A. G. A.mA. A.G. G*fU* G.mU.mU. G. G*fC* A* A* A. A.TEG-Chl 21168 2299 3403mC.mC.mU.mU.mU. 3404 P.mU.fC. A. A.fC.fU. A. 53% mC.mU. A. G. A.mA. A.G. G*mU* G.mU.mU. G. G*mC* A* A* A. A.TEG-Chl 21169 2299 3405mC.mC.mU.mU.mU. 3406 P.mU.fC. A. A.fC.fU. A. 47% mC.mU. A. G.mA. A.A.mG. G*fU* G.mU.mU. G. G*fC* A* A* A. A.TEG-Chl 21170 2299 3407mC.mC.mU.mU.mU. 3408 P.mU.fC. A. A.fC.fU. A. 70% mC.mU. A. G.mA. A.A.mG. G.mU.mU. G. G*mU* G*mC* A* A* A.TEG-Chl A. 21171 2299 3409mC.mC.mU.mU.mU. 3410 P.mU.fC. A. A.fC.fU. A. 65% mC.mU. A. G. A.mA. A.G. G*mU* G.mU.mU. G. G*mC* A*mA* A. A.TEG-Chl 21172 2299 3411mC.mC.mU.mU.mU. 3412 P.mU.fC. A. A.fC.fU. A. 43% mC.mU. A. G. A.mA. A.G. G*mU* G.mU.mU. G. G*mC*mA*mA* A. A.TEG-Chl 21173 2299 3413mC.mC.mU.mU.mU. 3414 P.mU.fC. A. A.fC.fU. A. 52% mC.mU. A. G. A.mA. A.G.mU.mU. G. G.mG*mU*mG*mC* A.TEG-Chl mA*mA* A. 21174 2299 3415mC.mC.mU.mU.mU. 3416 P.mU.fC. A. A.fC.fU. A. 47% mC.mU. A. G. A.mA. A.G. G.mU.mU. G. G*mU*mG*mC*mA* A.TEG-Chl mA* A. 21175 2299 3417mC.mC.mU.mU.mU. 3418 P.mU.fC. A. A.fC.fU. A. 35% mC.mU. A. G. A.mA. A.G. G.mU.mU. G. G*fU*mG*fC*mA*mA* A.TEG-Chl A. 21176 2299 3419mC.mC.mU.mU.mU. 3420 P.mU.fC. A. A.fC.fU. A. 50% mC.mU. A. G.mA. A.A.mG. G.mU.mU. G. G*fU*mG*fC*mA*mA* A.TEG-Chl A. 21177 2299 3421mC.mC.mU.mU.mU. 3422 P.mU.fC. A. A.fC.fU. A. 37% mC.mU. A. G. A. A. A.G. G*fU* G.mU.mU*mG*mA. G*fC* A* A* A. TEG-Chl 21178 2299 3423mC*mC*mU.mU. 3424 P.mU.fC. A. A.fC.fU. A. 36% mU.mC.mU. A. G. A. A. A.G. G*fU* G.mU.mU*mG*mA. G*fC* A* A* A. TEG-Chl 21179 2299 3425mC*mC*mU.mU. 3426 P.mU.fC. A. A.fC.fU. A. 35% mU.mC.mU.mA.mG. G. A. A.A. G. G*fU* mU.mU*mG*mA. G*fC* A* A* A. TEG-Chl 21203 2296 3427 G.mC.3428 P.mU.fC.fU. A. G. 40% A.mC.mC.mU.mU. A.mA. A. G. G.fU. mU.mC.mU.G.mC* A* A* A*mC* A*mG*mA.TEG- A* U. Chl 21204 2296 3429 G.mC. 3430P.mU.fC.fU. A. G.mA. 28% A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU.A* A* A*mC* A* U. A*mG*mA.TEG- Chl 21205 2296 3431 G.mC. 3432P.mU.fC.fU. A. G.mA. 51% A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU.A*mA* A*mC* A* U. A*mG*mA.TEG- Chl 21206 2296 3433 mG*mC* 3434P.mU.fC.fU. A. G. 46% A.mC.mC.mU.mU. A.mA. A. G. G.fU. mU.mC.mU. G.mC*A* A* A*mC* A*mG*mA.TEG- A* U. Chl 21207 2296 3435 mG*mC* 3436P.mU.fC.fU. A. G.mA. 29% A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU.A* A* A*mC* A* U. A*mG*mA.TEG- Chl 21208 2296 3437 mG*mC* 3438P.mU.fC.fU. A. G.mA. 72% A.mC.mC.mU.mU. A.mA. G. G.fU. G.mC* mU.mC.mU.A*mA* A*mC* A* U. A*mG*mA.TEG- Chl 21209 2296 3439 mG*mC*mA.mC. 3440P.mU.fC.fU. A. G. 89% mC.mU.mU.mU.mC. A.mA. A. G. G.fU. mU.mA*mG*mA.G.mC* A* A* A*mC* TEG-Chl A* U. 21210 2296 3441 mG*mC*mA.mC. 3442P.mU.fC.fU. A. G.mA. 65% mC.mU.mU.mU.mC. A.mA. G. G.fU. G.mC*mU.mA*mG*mA. A* A* A*mC* A* U. TEG-Chl 21211 2296 3443 mG*mC*mA.mC. 3444P.mU.fC.fU. A. G.mA. 90% mC.mU.mU.mU.mC. A.mA. G. G.fU. G.mC*mU.mA*mG*mA. A*mA* A*mC* A* U. TEG-Chl 21212 2295 3445 mU.mU. G.mC. 3446P.mU.fU. A. G. A.mA. 60% A.mC.mC.mU.mU. A. G. G.fU. mU.mC.mU*mA*G.fC.mA.mA*mA*fC* mA.TEG-Chl mA*mA*mG* G. 21213 2295 3447 mU.mU. G.mC.3448 P.mU.fU. A. G. A.mA. 63% A.mC.mC.mU.mU. A. G. G.fU. G.fC.mU.mC.mU*mA* A.mA*mA*mC*mA*mA* mA.TEG-Chl mG* G. 21214 2295 3449 mU.mU.G.mC. 3450 P.mU.fU. A. G. A.mA. 52% A.mC.mC.mU.mU. A. G. G.fU. G.fC. A.A* mU.mC.mU*mA* A*fC* A*mA*mG* G. mA.TEG-Chl 21215 2295 3451 mU.mU.G.mC. 3452 P.mU.fU. A. G. A.mA. 45% A.mC.mC.mU.mU. A. G. G.fU. G.fC.mA.mU.mC.mU*mA* A*mA*fC* mA.TEG-Chl A*mA*mG* G. 21216 2295 3453 mU*mU*G.mC. 3454 P.mU.fU. A. G. A.mA. 65% A.mC.mC.mU.mU. A. G. G.fU.mU.mC.mU*mA* G.fC.mA.mA*mA*fC* mA.TEG-Chl mA*mA*mG* G. 21217 2295 3455mU*mU* G.mC. 3456 P.mU.fU. A. G. A.mA. 69% A.mC.mC.mU.mU. A. G. G.fU.G.fC. mU.mC.mU*mA* A.mA*mA*mC*mA*mA* mA.TEG-Chl mG* G. 21218 2295 3457mU*mU* G.mC. 3458 P.mU.fU. A. G. A.mA. 62% A.mC.mC.mU.mU. A. G. G.fU.G.fC. A. A* mU.mC.mU*mA* A*fC* A*mA*mG* G. mA.TEG-Chl 21219 2295 3459mU*mU* G.mC. 3460 P.mU.fU. A. G. A.mA. 54% A.mC.mC.mU.mU. A. G. G.fU.G.fC.mA. mU.mC.mU*mA* A*mA*fC* mA.TEG-Chl A*mA*mG* G. 21220 2295 3461mU.mU.mG.mC.mA. 3462 P.mU.fU. A. G. A.mA. 52% mC.mC.mU.mU. A. G. G.fU.mU.mC.mU*mA* G.fC.mA.mA*mA*fC* mA.TEG-Chl mA*mA*mG* G. 21221 2295 3463mU.mU.mG.mC.mA. 3464 P.mU.fU. A. G. A.mA. 53% mC.mC.mU.mU. A. G. G.fU.G.fC. mU.mC.mU*mA* A.mA*mA*mC*mA*mA* mA.TEG-Chl mG* G. 21222 2295 3465mU.mU.mG.mC.mA. 3466 P.mU.fU. A. G. A.mA. 43% mC.mC.mU.mU. A. G. G.fU.G.fC. A. A* mU.mC.mU*mA* A*fC* A*mA*mG* G. mA.TEG-Chl 21223 2295 3467mU.mU.mG.mC.mA. 3468 P.mU.fU. A. G. A.mA. 43% mC.mC.mU.mU. A. G. G.fU.G.fC.mA. mU.mC.mU*mA* A*mA*fC* mA.TEG-Chl A*mA*mG* G. 21224 2299 3469mC.mC.mU.mU.mU. 3470 P.mU.fC. A. A.fC.fU. A. 60% mC.mU. A. G. A.mA. A.G. G.mU.mU*mG*mA. G*fU*mG*fC*mA*mA* TEG-Chl A. 21225 2299 3471mC*mC*mU.mU. 3472 P.mU.fC. A. A.fC.fU. A. 67% mU.mC.mU. A. G. A.mA. A.G. G.mU.mU*mG*mA. G*fU*mG*fC*mA*mA* TEG-Chl A. 21226 2299 3473mC*mC*mU.mU. 3474 P.mU.fC. A. A.fC.fU. A. 66% mU.mC.mU.mA.mG. G. A.mA.A. G. mU.mU*mG*mA. G*fU*mG*fC*mA*mA* TEG-Chl A. 21227 2296 3475 G.mC.3476 P.mU.fC.fU. A. G.mA. 49% A.mC.mC.mU.mU. A.mA. G. G.fU. mU.mC.mU.G.fC*mA*mA*mA*fC* A*mG*mA.TEG- mA* U. Chl 20584 2296 3477 G.mC. 3478P.mU.fC.fU. A. G. A. A. 70% A.mC.mC.mU.mU. A. G. G.mU. G.mC* A*mU.mC.mU. A. G. A* A*mC* A* U. A.Chl-TEG 20585 2296 3479 G.mC. 3480P.mU.fC.fU. A. G. A. A. 15% A.mC.mC.mU.mU. A. G. G.fU. G.mC* A*mU.mC.mU. A. G. A* A*mC* A* U. A.Chl-TEG 20586 2296 3481 G.mC. 3482P.mU. C. U. A. G. A. A. 30% A.mC.mC.mU.mU. A. G. G.mU. G.mC* A*mU.mC.mU. A. G. A* A*mC* A* U. A.Chl-TEG 20587 2296 3483 G.mC. 3484P.mU.fC.fU. A. G. A. A. 32% A.mC.mC.mU.mU. A. G. G.fU. mU.mC.mU. A. G.G.fC*mA*mA*mA*fC* A.Chl-TEG mA* U. 20616 2275 3485 G.mU. G. 3486 P.mU.22% A.mC.mC. A. A. A. A.fC.fU.fU.fU.fU. G. A. G.mU. A.Chl- G.fU.mC.A.mC* TEG A*mC*mU*mC*mU* C. 20617 2275 3487 G.mU. G. 3488 P.mU. 18%A.mC.mC. A. A. A. A.fC.fU.fU.fU.fU. G. A. G.mU. A.Chl- G.fU.fC. A.mC*TEG A*fC*mU*fC*mU* C. 20618 2275 3489 G.mU. G. 3490 P.mU. A. C. U. U. U.U. 36% A.mC.mC. A. A. A. G. G. U.mC. A.mC* A. G.mU. A.Chl-A*mC*mU*mC*mU* TEG C. 20619 2275 3491 G.mU. G. 3492 P.mU. 28% A.mC.mC.A. A. A. A.fC.fU.fU.fU.fU. G. A. G.mU. A.Chl- G.fU.fC. TEGA.mC*mA*mC*mU*mC* mU* C. 21381 2275 3493 G.mU. G. 3494 P.mU. 28%A.mC.mC. A. A. A. A.fC.fU.fU.fU.fU. G. A. G.fU.mC. A.mC* G*mU*mA.TEG-A*mC*mU*mC*mU* Chl C. 21382 2275 3495 G.mU. G. 3496 P.mU. 28% A.mC.mC.A. A. A. A.fC.fU.fU.fU.fU. G. A. G.fU.fC. A.mC* G*mU*mA.TEG-A*fC*mU*fC*mU* C. Chl 21383 2275 3497 mG*mU*mG.mA. 3498 P.mU. 43%mC.mC.mA.mA.mA. A.fC.fU.fU.fU.fU. G. mA.mG*mU*mA. G.fU.mC. A.mC* TEG-ChlA*mC*mU*mC*mU* C. 21384 2275 3499 mG*mU*mG.mA. 3500 P.mU. 50%mC.mC.mA.mA.mA. A.fC.fU.fU.fU.fU. G. mA.mG*mU*mA. G.fU.fC. A.mC* TEG-ChlA*fC*mU*fC*mU* C. 20392 2275 3501 G.mU. G. 3502 P.mU. 28% A.mC.mC. A. A.A. A.fC.fU.fU.fU.fU. G. A. G.mU. A.TEG- G.fU.fC. A.fC* ChlA*fC*fU*fC*fU* C. 20393 2296 3503 G.mC. 3504 P.mU.fC.fU. A. G. A. A. 35%A.mC.mC.mU.mU. A. G. G.fU. G.fC* A* mU.mC.mU. A. G. A* A*fC* A* U.A.TEG-Chl 21429 2275 3505 G.mU. G. 3506 P.mU. 36% A.mC.mC. A. A. A.A.fC.fU.fU.fU.fU. G. A. G.fU.fC. A.mC* G*mU*mA.Teg- A*fC*mU*fC*mU* C.Chl 21430 2275 3507 G.mU. G. 3508 P.mU. 31% A.mC.mC. A.A.fC.fU.fU.fU.fU. G. A.mA. A. G.fU.mC. A.mC* G*mU*mA.Teg- A*mC*mU*mC*mU*Chl C.

TABLE 21 Inhibition of gene expression with TGFB2 sd-rxRNA sequences(Accession Number: NM_001135599.1) % remaining Oligo Start SEQ SEQ IDexpression (1 Number Site ID NO Sense sequence NO Antisense sequence uM,A549) 14408 1324 3509 G. 3510 P.mU.fC. G. A. A. G. G. 94% G.mC.mU.mC.mU.A. G. A. G.mC.mC* mC.mC.mU.mU.mC. A*mU*mU*mC* G* C. G. A.Chl 14409 13743511 G. A.mC. A. G. G. A. 3512 P.mC.fC. A. G. n/a A.mC.mC.mU. G.G.fU.fU.fC.fC.fU. G.Chl G.mU.mC*mU*mU*mU* A*mU* G. 14410 946 3513 mC.mC.A. A. G. G. 3514 P.mU. A. A. 90% A. G. A.fC.fC.fU.fC.fC.fU.mU.G.mU.mU.mU. G. G*mC* G*mU* A* A.Chl G* U. 14411 849 3515 A.mU.mU.mU.mC.3516 P.mU. G.fU. A. G. A.fU. 72% mC. A.mU.mC.mU. G. G. A. A. A.mU*mC*A.mC. A.Chl A*mC*mC*mU* C. 14412 852 3517 mU.mC.mC. 3518 P.mU. G.fU.fU.G.fU. 76% A.mU.mC.mU. A. G. A.fU. G. G. A* A* A.mC. A. A.mC. A*mU*mC* A*C. A.Chl 14413 850 3519 mU.mU.mU.mC.mC. 3520 P.mU.fU. G.fU. A. G. 98%A.mU.mC.mU. A.fU. G. G. A. A. A.mC. A. A.Chl A*mU*mC* A*mC*mC* U. 14414944 3521 mC. G.mC.mC. A. A. 3522 P.mA. 100% G. G. A. G.A.fC.fC.fU.fC.fC.fU.fU. G.mU.mU.Chl G. G.mC. G*mU* A* G*mU* A* C. 144151513 3523 G.mU. G. G.mU. G. 3524 P.mU.fU.fC.fU. G. n/a A.mU.mC. A. G. A.A.fU.fC. A.fC.mC. A.Chl A.mC*mU* G* G*mU* A* U. 14416 1572 3525mC.mU.mC.mC.mU. 3526 P.mA.fC. A.fU.fU. A. 100% G.mC.mU. A. G.fC. A. G.G. A. G* A.mU. G.mU.Chl A*mU* G*mU* G* G. 14417 1497 3527A.mC.mC.mU.mC.mC. 3528 P.mU. A.fU. A.fU. 73% A.mC. A.mU. G.fU. G. G. A.G. A.mU. A.Chl G.mU* G*mC*mC* A*mU* C. 14418 1533 3529 A. A. G.mU.mC.mC.3530 P.mU.fC.fC.fU. A. G.fU. 98% A.mC.mU. A. G. G. G. G. A.ChlA.mC.mU.mU*mU* A*mU* A* G* U. 14419 1514 3531 mU. G. G.mU. G. 3532P.mU.fU.fU.fC.fU. G. 86% A.mU.mC. A. G. A. A.fU.fC. A.mC.mC. A. A.ChlA*mC*mU* G* G*mU* A. 14420 1534 3533 A. G.mU.mC.mC. 3534P.mU.fU.fC.fC.fU. A. 99% A.mC.mU. A. G. G. G.fU. G. G. A. A.ChlA.mC.mU*mU*mU* A*mU* A* G. 14421 943 3535 A.mC. G.mC.mC. A. 3536P.mA.fC.fC.fU.fC.fC.fU. 41% A. G. G. A. G. fU. G. G.mC. G.mU* G.mU.ChlA* G*mU* A*mC* U. 18570 2445 3537 mU. A.mU.mU.mU. 3538 P.mU. A.fC. A.fC.A. 79% A.mU.mU. G.mU. A.fU. A. A. A.fU. A* G.mU. A.Chl A*fC*fU*fC* A* C.18571 2445 3539 mU.mU. 3540 P.mU. A.fC. A.fC. A. 75% A.mU.mU.mU. A.fU.A. A. A.fU. A* A.mU.mU. G.mU. A*fC*fU*fC* A* C. G.mU. A.Chl 18572 20833541 A.mU. C. A. G.mU. 3542 P.mU.fU.fU.fU. A. 47% G.mU.mU. A. A. A.A.fC. A.fC.fU. G. A.fU* A.Chl G* A* A*fC*fC* A. 18573 2083 3543 mC.A.mU.mC. A. 3544 P.mU.fU.fU.fU. A. 17% G.mU. G.mU.mU. A.fC. A.fC.fU. G.A.fU* A. A. A. A.Chl G* A* A*fC*fC* A. 18574 2544 3545 A.mU. G. 3546P.mU.fU.fC.fC.fU.fU. 59% G.mC.mU.mU. A. A. A. A. G.fC.fC. A. G. G. A.A.Chl U*fC*fC* A*fU* G* A. 18575 2544 3547 G. A.mU. G. 3548P.mU.fU.fC.fC.fU.fU. 141% G.mC.mU.mU. A. A. A. A. G.fC.fC. A. G. G. A.A.Chl U*fC*fC* A*fU* G* A. 18576 2137 3549 mU.mU. G.mU. 3550 P.mU. A.A.fC. A. G. A. 77% G.mU.mU.mC.mU. A.fC. A.fC. A. A* G.mU.mU. A.ChlA*fC*fU*fU*fC* C. 18577 2137 3551 mU.mU.mU. G.mU. 3552 P.mU. A. A.fC. A.G. A. 59% G.mU.mU.mC.mU. A.fC. A.fC. A. A* G.mU.mU. A.Chl A*fC*fU*fU*fC*C. 18578 2520 3553 A. A. A.mU. 3554 P.mU. G. G.fC. A. A. A. 75%A.mC.mU.mU.mU. G.fU. A.fU.fU.fU* G* G.mC.mC. A.Chl G*fU*fC*fU* C. 185792520 3555 mC. A. A. A.mU. 3556 P.mU. G. G.fC. A. A. A. 55%A.mC.mU.mU.mU. G.fU. A.fU.fU.fU* G* G.mC.mC. A.Chl G*fU*fC*fU* C. 185803183 3557 mC.mU.mU. G.mC. 3558 P.mU.fU.fU. G.fU. A. 84% A.mC.mU. A.mC.A. G.fU. G.fC. A. A. A. A.Chl G*fU*fC* A* A* A* C. 18581 3183 3559A.mC.mU.mU. 3560 P.mU.fU.fU. G.fU. A. 80% G.mC. A.mC.mU. G.fU. G.fC. A.A. A.mC. A. A. A.Chl G*fU*fC* A* A* A* C. 18582 2267 3561 G. A. 3562P.mU. A.fC.fU. A. A.fU. 82% A.mU.mU.mU. A. A. A.mU.mU. A. G.mU.A.fU.fU.fC*fU*fU*fC*fC* A.Chl A* G. 18583 2267 3563 A. G. A. 3564 P.mU.A.fC.fU. A. A.fU. 67% A.mU.mU.mU. A. A. A.mU.mU. A. G.mU.A.fU.fU.fC*fU*fU*fC*fC* A.Chl A* G. 18584 3184 3565 mU.mU. G.mC. 3566P.mU.fU.fU.fU. G.fU. 77% A.mC.mU. A.mC. A. A. G.fU. G.fC. A. A* A. A.A.Chl G*fU*fC* A* A* A. 18585 3184 3567 mC.mU.mU. G.mC. 3568P.mU.fU.fU.fU. G.fU. 59% A.mC.mU. A.mC. A. A. G.fU. G.fC. A. A* A. A.A.Chl G*fU*fC* A* A* A. 18586 2493 3569 A.mU. A. A. A. 3570 P.mU.fC.A.fC.fC.fU. 84% A.mC. A. G. G.mU. G.fU.fU.fU.fU. G. A.ChlA.fU*fU*fU*fU*fC*fC* A. 18587 2493 3571 A. A.mU. A. A. A. 3572 P.mU.fC.A.fC.fC.fU. 70% A.mC. A. G. G.mU. G.fU.fU.fU.fU. G. A.ChlA.fU*fU*fU*fU*fC*fC* A. 18588 2297 3573 G. A.mC. A. A.mC. 3574 P.mU.G.fU.fU. 40% A. A.mC. A. A.mC. G.fU.fU. G.fU.fU. A.Chl G.fU.fC* G*fU*fU*G*fU* U. 18589 2046 3575 A.mU. G. 3576 P.mU.fU. G.fU.fU. 39% C.mU.mU.G.mU. A. A.fC. A. A. G.fC. A.mC. A. A.Chl A.fU*fC* A*fU*fC* G* U. 185902531 3577 mC. A. G. A. A. 3578 P.mU.fC. A.fU. G. A. 56% A.mC.mU.mC.G.fU.fU.fU.fC.fU. G* A.mU. G. A.Chl G*fC* A* A* A* G. 18591 2389 3579G.mU. A.mU.mU. 3580 P.mU. G.fC. A.fU. A. 64% G.mC.mU. A.mU. G.fC. A.A.fU. A.fC* A* G.mC. A.Chl G* A* A* A* A. 18592 2530 3581 mC.mC. A. G.A. A. 3582 P.mU. A.fU. G. A. 44% A.mC.mU.mC. G.fU.fU.fU.fC.fU. G. A.mU.A.Chl G*fC* A* A* A* G* U. 18593 2562 3583 A.mC.mU.mC. A. A. 3584 P.mU.G.fC.fU.fC. 87% A.mC. G. A. G.mC. G.fU.fU.fU. G. A. A.Chl G.fU*fU*fC* A*A* G* U. 18594 2623 3585 A.mU. A.mU. G. 3586 P.mU.fU.fC.fU.fC. G. 69%A.mC.mC. G. A. G. G.fU.fC. A.fU. A.fU* A. A.Chl A* A*fU* A* A* C. 185952032 3587 mC. G. A.mC. G. 3588 P.mU.fU.fC. G.fU.fU. 55% A.mC. A. A.mC.G. G.fU.fC. G.fU.fC. A. A.Chl G*fU*fC* A*fU*fC* A. 18596 2809 3589 G.mU.A. A. 3590 P.mU.fU.fC. A.fC.fU. G. 58% A.mC.mC. A. G.mU. G.fU.fU.fU.A.fC*fU* G. A. A.Chl A* A* A*fC* U. 18597 2798 3591 mU.mU. G.mU.mC. 3592P.mU.fC.fU. A. A. 38% A. G.mU.mU.mU. A. A.fC.fU. G. A.fC. A. A* G. A.ChlA* G* A* A*fC* C. 18598 2081 3593 mU.mC. A.mU.mC. 3594 P.mU.fU. A. A.fC.25% A. G.mU. A.fC.fU. G. A.fU. G. A* G.mU.mU. A. A.Chl A*fC*fC* A* A* G.18599 2561 3595 A. A.mC.mU.mC. A. 3596 P.mU.fC.fU.fC. 57% A. A.mC. G. A.G. G.fU.fU.fU. G. A. A.Chl G.fU.fU*fC* A* A* G*fU* U. 18600 2296 3597mC. G. A.mC. A. 3598 P.mU.fU.fU. G.fU.fU. 69% A.mC. A. A.mC. A. G.fU.fU.G.fU.fC. A. A.Chl G*fU*fU* G*fU*fU* C. 18601 2034 3599 A.mC. G. A.mC. A.3600 P.mU.fC. A.fU.fC. 22% A.mC. G. A.mU. G. G.fU.fU. G.fU.fC. A.ChlG.fU*fC* G*fU*fC* A*fU. 18602 2681 3601 G.mC.mU. 3602P.mU.fU.fC.fC.fU.fU. 43% G.mC.mC.mU. A. A. A. G. G.fC. A. G.fC*fU* G. G.A. A.Chl G* A*fU* A* C. 18603 2190 3603 A.mU.mU.mC.mU. 3604 P.mU. G. A.A. A.fU. 128% A.mC. G.fU. A. G. A. A.fU* A* A.mU.mU.mU.mC. A* G* G*fC*C. A.Chl 20604 2083 3605 mC. A.mU.mC. A. 3606 P.mU.fU.fU.fU. A. 19%G.mU. G.mU.mU. A.fC. A.fC.fU. G. A. A. A. A.Chl A.mU* G* A* A*mC*mC* A.20605 2083 3607 mC. A.mU.mC. A. 3608 P.mU.fU.fU.fU. A. 20% G.mU.G.mU.mU. A.fC. A.fC.fU. G. A. A. A. A.Chl A.mU* G* A* A*fC*mC* A. 206062083 3609 mC. A.mU.mC. A. 3610 P.mU. U. U. U. A. A. C. 82% G.mU.G.mU.mU. A. C. U. G. A.mU* G* A. A. A. A.Chl A* A*mC*mC* A. 20607 20833611 mC. A.mU.mC. A. 3612 P.mU.fU.fU.fU. A. 59% G.mU. G.mU.mU. A.fC.A.fC.fU. G. A. A. A. A.Chl A.fU*mG*mA*mA*fC* fC* A. 21722 2081 3613mU.mC. A.mU.mC. 3614 P.mU.fU. A. A.fC. 34% A. G.mU. A.fC.fU. G. A.fU. G.A* G.mU.mU. A. A.Chl A*mC*mC* A* A* G. 21723 2081 3615 mU.mC. A.mU.mC.3616 P.mU.fU. A. A.fC. 53% A. G.mU. A.fC.fU. G. A.fU. G.mU.mU. A. A.ChlG.mA*mA*mC*mC*mA* mA* G. 21724 2081 3617 mU.mC. A.mU.mC. 3618 P.mU.fU.A. A.fC. 48% A. G.mU. A.fC.fU. G. A.mU. G.mU.mU. A. A.ChlG.mA*mA*mC*mC*mA* mA* G. 21725 2081 3619 mU.mC. A.mU.mC. 3620 P.mU.fU.A. A.fC. 45% A. G.mU. A.fC.fU. G. A.fU. G. A* G.mU.mU. A. A.ChlA*fC*fC*mA*mA* G. 21726 2081 3621 mU.mC. A.mU.mC. 3622 P.mU.fU. A. A.fC.54% A. G.mU. A.fC.fU. G. A.fU. G.mU.mU. A. A.Chl G.mA*mA*fC*fC*mA* mA*G. 21727 2081 3623 mU.mC. A.mU.mC. 3624 P.mU.fU.A. A.fC. 29% A. G.mU.A.fC.fU. G. A.fU. G. A* G.mU.mU*mA*mA. A*fC*fC* A* A* G. TEG-Chl 217282081 3625 mU*mC* A.mU.mC. 3626 P.mU.fU. A. A.fC. 27% A. G.mU. A.fC.fU.G. A.fU. G. A* G.mU.mU*mA*mA. A*fC*fC* A* A* G. TEG-Chl 21729 2081 3627mU*mC*mA.mU.mC. 3628 P.mU.fU. A. A.fC. 30% mA.mG.mU.mG. A.fC.fU. G.A.fU. G. A* mU.mU*mA*mA.TEG- A*fC*fC* A* A* G. Chl 21375 2081 3629mU.mC. A.mU.mC. 3630 P.mU.fU. A. A.fC. 29% A. G.mU. A.fC.fU. G. A.fU. G.A* G.mU.mU*mA*mA. A*mC*mC* A* A* G. TEG-Chl 21376 2081 3631 mU.mC.A.mU.mC. 3632 P.mU.fU. A. A.fC. 30% A. G.mU. A.fC.fU. G. A.fU. G. A*G.mU.mU*mA*mA. A*fC*fC*mA*mA* G. TEG-Chl 21377 2081 3633 mU.mC. A.mU.mC.3634 P.mU.fU. A. A.fC. 37% A. G.mU. A.fC.fU. G. A.fU. G.mU.mU*mA*mA.G.mA*mA*fC*fC*mA* TEG-Chl mA* G. 21378 2081 3635 mU*mC*mA.mU.mC. 3636P.mU.fU. A. A.fC. 32% mA.mG.mU.mG. A.fC.fU. G. A.fU. G. A*mU.mU*mA*mA.TEG- A*mC*mC* A* A* G. Chl 21379 2081 3637 mU*mC*mA.mU.mC.3638 P.mU.fU. A. A.fC. 31% mA.mG.mU.mG. A.fC.fU. G. A.fU. G. A*mU.mU*mA*mA.TEG- A*fC*fC*mA*mA* G. Chl 21380 2081 3639 mU*mC*mA.mU.mC.3640 P.mU.fU. A. A.fC. 39% mA.mG.mU.mG. A.fC.fU. G. A.fU.mU.mU*mA*mA.TEG- G.mA*mA*fC*fC*mA* Chl mA* G.

TABLE 22 Inhibition of gene expression with TGFB1 sd-rxRNA sequences(Accession Number: NM_000660.3) % remaining Oligo Start SEQ ID SEQ IDexpression Number Site NO Sense sequence NO Antisense sequence (1 uMA549) 14394 1194 3641 G.mC.mU. A. A.mU. 3642 P.mU.fU.fC.fC. A.fC.fC. 24%G. G.mU. G. G. A. A.fU.fU. A. G.mC* A.Chl A*mC* G*mC* G* G. 14395 20063643 mU. G. A.mU.mC. 3644 P.mG. A. G.fC. G.fC. 79% G.mU. G.mC. A.fC. G.A.mU.mC. G.mC.mU.mC.Chl A*mU* G*mU*mU* G* G. 14396 1389 3645 mC. A. 3646P.mU.fC. G.fC.fC. A. G. 77% A.mU.mU.mC.mC.mU. G. A. A.mU.mU. G. G.mC. G.A.Chl G*mU*mU* G*mC*mU* G. 14397 1787 3647 A. G.mU. G. G. 3648 P.mU.fC.G.fU. G. G. n/a A.mU.mC.mC. A.mC. A.fU.fC.fC. G. A.Chl A.mC.mU*mU*mC*mC*A* G* C. 14398 1867 3649 mU. A.mC. A. G.mC. 3650 P.mG. G. A.fC.fC.fU.fU.82% A. A. G. G.fC.fU. G.mU. G.mU.mC.mC.Chl A*mC*mU* G*mC* G* U. 143992002 3651 A. A.mC. A.mU. G. 3652 P.mG.fC. A.fC. G. n/a A.mU.mC. G.mU.A.fU.fC. A.fU. G.mC.Chl G.mU.mU* G* G* A*mC* A* G. 14400 2003 3653 A.mC.A.mU. G. 3654 P.mC. G.fC. A.fC. G. n/a A.mU.mC. G.mU. A.fU.fC. A.mU.G.mC. G.Chl G.mU*mU* G* G* A*mC* A. 14401 1869 3655 mC. A. G.mC. A. A.G. 3656 P.mC. A. G. G. 82% G.mU.mC.mC.mU. A.fC.fC.fU.fU. G.Chl G.mC.mU.G*mU* A*mC*mU* G* C. 14402 2000 3657 mC.mC. A. A.mC. 3658 P.mA.fC. G.A.fU.fC. 66% A.mU. G. A.mU.mC. A.fU. G.fU.mU. G. G* G.mU.Chl A*mC* A*G*mC* U. 14403 986 3659 A. G.mC. G. G. A. A. 3660 P.mA.fU. G.fC. 78%G.mC. G.mC. G.fC.fU.fU.fC.fC. A.mU.Chl G.mC.mU*mU*mC* A*mC*mC* A. 14404995 3661 G.mC. A.mU.mC. G. 3662 P.mA.fU. G. 79% A. G. G.mC.mC.G.fC.fC.fU.fC. G. A.mU. A.mU.Chl G.mC* G*mC*mU*mU*mC* C. 14405 963 3663G. A.mC.mU. 3664 P.mC. A.fU. G.fU.fC. G. 80% A.mU.mC. G. A.mC. A.fU. A.A.mU. G.Chl G.mU.mC*mU*mU* G*mC* A* G. 14406 955 3665 A.mC.mC.mU. G.mC.3666 P.mU. A. G.fU.fC.fU.fU. 88% A. A. G. A.mC.mU. G.fC. A. G. G.mU* G*A.Chl G* A*mU* A* G. 14407 1721 3667 G.mC.mU.mC.mC. 3668P.mU.fU.fC.fU.fC.fC. n/a A.mC. G. G. A. G. A. G.fU. G. G. A. A.ChlG.mC*mU* G* A* A* G* C. 18454 1246 3669 mC. A.mC. A. G.mC. 3670 P.mU.A.fU. A.fU. A.fU. 58% A.mU. A.mU. A.mU. G.fC.fU. G.fU. G*fU* A.Chl G*fU*A*fC* U. 18455 1248 3671 mC. A. G.mC. A.mU. 3672 P.mU. A.fU. A.fU. A.fU.87% A.mU. A.mU. A.mU. A.fU. G.fC.fU. G*fU* A.Chl G*fU* G*fU* A. 184561755 3673 G.mU. A.mC. 3674 P.mU. A. A. G.fU.fC. A. 107%  A.mU.mU. G.A.fU. G.fU. A.fC* A* A.mC.mU.mU. A.Chl G*fC*fU* G* C. 18457 1755 3675mU. G.mU. A.mC. 3676 P.mU. A. A. G.fU.fC. A. 77% A.mU.mU. G. A.fU. G.fU.A.fC* A* A.mC.mU.mU. A.Chl G*fC*fU* G* C. 18458 1708 3677 A. A.mC.mU.3678 P.mU. G. A. A. G.fC. A. 75% A.mU.mU. A.fU. A. G.fU.fU* G*G.mC.mU.mU.mC. G*fU* G*fU* C. A.Chl 18459 1708 3679 mC. A. A.mC.mU. 3680P.mU. G. A. A. G.fC. A. 73% A.mU.mU. A.fU. A. G.fU.fU* G* G.mC.mU.mU.mC.G*fU* G*fU* C. A.Chl 18460 1250 3681 G.mC. A.mU. A.mU. 3682 P.mU. A.fC.A.fU. A.fU. n/a A.mU. A.mU. G.mU. A.fU. A.fU. G.fC*fU* A.Chl G*fU* G*fU*G. 18461 1754 3683 mU. G.mU. A.mC. 3684 P.mU. A. G.fU.fC. A. 91%A.mU.mU. G. A.fU. G.fU. A.fC. A* A.mC.mU. A.Chl G*fC*fU* G*fC* C. 184621754 3685 mC.mU. G.mU. 3686 P.mU. A. G.fU.fC. A. 92% A.mC. A.mU.mU. G.A.fU. G.fU. A.fC. A* A.mC.mU. A.Chl G*fC*fU* G*fC* C. 18463 1249 3687 A.G.mC. A.mU. 3688 P.mU.fC. A.fU. A.fU. n/a A.mU. A.mU. A.mU. A.fU. A.fU.G.fC.fU* G. A.Chl G*fU* G*fU* G* U. 18464 1383 3689 mC. A. G.mC. A. 3690P.mU. G. A. A.fU.fU. 77% A.mC. A. G.fU.fU. G.fC.fU. G*fU* A.mU.mU.mC.A.Chl A*fU*fU*fU* C. 18465 1251 3691 mC. A.mU. A.mU. 3692 P.mU. A. A.fC.A.fU. 84% A.mU. A.mU. A.fU. A.fU. A.fU. G.mU.mU. A.Chl G*fC*fU* G*fU* G*U. 18466 1713 3693 mU.mU. 3694 P.mU. G. A. G.fC.fU. G. n/aG.mC.mU.mU.mC. A. A. A. G.fC. A. A*fU* A* G.mC.mU.mC. A.Chl G*fU*fU* G.18467 1713 3695 A.mU.mU. 3696 P.mU. G. A. G.fC.fU. G. 83% G.mC.mU.mU.mC.A. A. A. G.fC. A. A*fU* A* G.mC.mU.mC. A.Chl G*fU*fU* G. 18468 1247 3697A.mC. A. G.mC. 3698 P.mU.fU. A.fU. A.fU. 96% A.mU. A.mU. A.mU. A.fU.G.fC.fU. G.fU* A. A.Chl G*fU* G*fU* A* C. 18469 1712 3699 A.mU.mU. 3700P.mU. A. G.fC.fU. G. A. 90% G.mC.mU.mU.mC. A. A. G.fC. A. A.fU* A*G.mC.mU. A.Chl G*fU*fU* G* G. 18470 1712 3701 mU. A.mU.mU. 3702 P.mU. A.G.fC.fU. G. A. 98% G.mC.mU.mU.mC. A. A. G.fC. A. A.fU* A* G.mC.mU. A.ChlG*fU*fU* G* G. 18471 1212 3703 mC. A. A. 3704 P.mU.fU. G.fC.fU.fU. G.n/a G.mU.mU.mC. A. A. A. A.fC.fU.fU. G*fU*fC* G.mC. A. A.Chl A*fU* A* G.18472 1222 3705 mC. A. G. A. G.mU. 3706 P.mU. G.fU. G.fU. G.fU. 45%A.mC. A.mC. A.mC. A.fC.fU.fC.fU. G* A.Chl C*fU*fU* G* A* A. 18473 12283707 A.mC. A.mC. A.mC. 3708 P.mU.fU. A.fU. G.fC.fU. 36% A. G.mC. A.mU.A. G.fU. G.fU. G.fU* A.Chl A*fC*fU*fC*fU* G. 18474 1233 3709 mC. A.G.mC. A.mU. 3710 P.mU. A.fU. A.fU. A.fU. 68% A.mU. A.mU. A.mU. A.fU.G.fC.fU. G*fU* A.Chl G*fU* G*fU* A. 18475 1218 3711 mU.mC. A. A. G.mC.3712 P.mU.fU. A.fC.fU.fC.fU. 64% A. G. A. G.mU. A. G.fC.fU.fU. G. A*A.Chl A*fC*fU*fU* G* U. 18476 1235 3713 A. G.mC. A.mU. 3714 P.mU.fC.A.fU. A.fU. 78% A.mU. A.mU. A.mU. A.fU. A.fU. G.fC.fU* G. A.Chl G*fU*G*fU* G* U. 18477 1225 3715 A. G. A. G.mU. A.mC. 3716 P.mU.fU. G.fU.G.fU. 92% A.mC. A.mC. A. A.Chl G.fU. A.fC.fU.fC.fU* G*fC*fU*fU* G* A.18478 1221 3717 A. A. G.mC. A. G. A. 3718 P.mU.fU. G.fU. 103%  G.mU.A.mC. A. A.Chl A.fC.fU.fC.fU. G.fC.fU.fU* G* A* A*fC*fU* U. 18479 12443719 mU.mU.mC. A. A.mC. 3720 P.mU.fU. G. A.fU. G.fU. 84% A.mC. A.mU.mC.A. G.fU.fU. G. A. A* G* A* A.Chl A*fC* A* U. 18480 1224 3721 A. G.mC. A.G. A. 3722 P.mU. G.fU. G.fU. 37% G.mU. A.mC. A.mC. A.fC.fU.fC.fU. A.ChlG.fC.fU*fU* G* A* A*fC* U. 18481 1242 3723 A.mU. A.mU. A.mU. 3724 P.mU.A. A. G. A. A.fC. 62% G.mU.mU.mC.mU.mU. A.fU. A.fU. A.fU* A*fU* A.ChlG*fC*fU* G. 18482 1213 3725 G. A.mC. A. A. 3726 P.mU.fC.fU.fU. G. A. 47%G.mU.mU.mC. A. A. A.fC.fU.fU. G.fU.fC* G. A.Chl A*fU* A* G* A* U. 184831760 3727 mU.mU. A. A. A. G. 3728 P.mU.fC.fU.fC.fC. 69% A.mU. G. G. A.G. A.fU.fC.fU.fU.fU. A. A.Chl A*fU* G* G* G* G* C. 18484 1211 3729mC.mU. A.mU. G. 3730 P.mU. A. A.fC.fU.fU. n/a A.mC. A. A. G.fU.fC. A.fU.A. G* G.mU.mU. A.Chl A*fU*fU*fU*fC* G. 19411 1212 3731 mC. A. A.mC. G.A. A. 3732 P.mU.fU. A. G. 52% A.mU.mC.mU. A. A.fU.fU.fU.fC. G.fU.fU.A.Chl G*fU* G* G* G*fU*fU. 19412 1222 3733 mU. A.mU. G. A.mC. 3734 P.mU.G. A. A.fC.fU.fU. 51% A. A. G.mU.mU.mC. G.fU.fC. A.fU. A* G* A.ChlA*fU*fU*fU*fC. 19413 1228 3735 A. A. G.mU.mU.mC. 3736 P.mU.fC.fU.G.fC.fU.fU. n/a A. A. G.mC. A. G. G. A. A.fC.fU.fU* A.Chl G*fU*fC* A*fU*A. 19414 1233 3737 mC. A. A. G.mC. A. G. 3738 P.mU. G.fU. 41% A. G.mU.A.mC. A.Chl A.fC.fU.fC.fU. G.fC.fU.fU. G* A* A*fC*fU*fU* G. 19415 12183739 A. A.mU.mC.mU. 3740 P.mU.fU.fU. G.fU.fC. 104%  A.mU. G. A.mC. A. A.A.fU. A. G. A.Chl A.fU.fU*fU*fC* G*fU*fU* G. 19416 1244 3741 mC. A.mC.A.mC. A. 3742 P.mU. A.fU. A.fU. 31% G.mC. A.mU. A.mU. G.fC.fU. G.fU.G.fU. A.Chl G*fU* A*fC*fU*fC*fU. 19417 655 3743 G. A. A. A.mU. 3744P.mU.fU.fU. G.fC.fU. n/a A.mU. A. G.mC. A. A. A.fU. A.fU.fU.fU.fC*fU*A.Chl G* G*fU* A* G. 19418 644 3745 G. A. 3746 P.mU.fC.fU. G. G.fU. A.n/a A.mC.mU.mC.mU. G. A. G.fU.fU.fC*fU* A.mC.mC. A. G. A.Chl A*fC* G*fU*G. 19419 819 3747 G.mC. A. A. A. G. 3748 P.mU.fC. A.fU.fU. n/a A.mU. A.A.mU. G. A.fU.fC.fU.fU.fU. A.Chl G.fC*fU* G*fU*fC* A* C. 19420 645 3749A. A.mC.mU.mC.mU. 3750 P.mU.fU.fC.fU. G. G.fU. n/a A.mC.mC. A. G. A. A.G. A. G.fU.fU*fC*fU* A.Chl A*fC* G* U. 19421 646 3751 A.mC.mU.mC.mU.3752 P.mU.fU.fU.fC.fU. G. n/a A.mC.mC. A. G. A. A. G.fU. A. G. A. A.ChlG.fU*fU*fC*fU* A*fC* G. 19422 816 3753 A.mC. A. G.mC. A. A. 3754P.mU.fU. n/a A. G. A.mU. A. A.Chl A.fU.fC.fU.fU.fU. G.fC.fU. G.fU*fC*A*fC* A* A* G. 19423 495 3755 mC. A. A.mU.mC.mU. 3756 P.mU.fU. G.fU.fC.A.fU. n/a A.mU. G. A.mC. A. A. G. A.fU.fU. G*fC* A.Chl G*fU*fU* G* U.19424 614 3757 A. G. A.mU.mU.mC. 3758 P.mU.fU. G. A.fC.fU.fU. n/a A. A.G.mU.mC. A. G. A. A.fU.fC.fU*fC*fU* A.Chl G*fC* A* G. 19425 627 3759mC.mU. G.mU. G. G. 3760 P.mU. G.fU.fU. n/a A. G.mC. A. A.mC.G.fC.fU.fC.fC. A.fC. A. A.Chl G*fU*fU* G* A*fC* U. 19426 814 3761 mU. G.A.mC. A. 3762 P.mU.fU.fC.fU.fU.fU. n/a G.mC. A. A. A. G. A. G.fC.fU.G.fU.fC. A*fC* A.Chl A* A* G* A* G. 19427 501 3763 A.mU. G. A.mC. A. A.3764 P.mU.fU. G. n/a A. A.mC.mC. A. A.Chl G.fU.fU.fU.fU. G.fU.fC. A.fU*A* G* A*fU*fU* G. 19428 613 3765 G. A. G. 3766 P.mU. G. A.fC.fU.fU. G.n/a A.mU.mU.mC. A. A. A. A.fU.fC.fU.fC*fU* G.mU.mC. A.Chl G*fC* A* G* G.21240 1244 3767 mC. A.mC. A.mC. A. 3768 P.mU. A.fU. A.fU. 0.875 G.mC.A.mU. A.mU. G.fC.fU. G.fU. G.fU. A.Chl G*mU* A*mC*mU*mC* U. 21241 12443769 mC. A.mC. A.mC. A. 3770 P.mU. A.fU. A.fU. 0.88 G.mC. A.mU. A.mU.G.fC.fU. G.fU. G.fU. A.Chl G*mU*mA*mC*mU*mC* U. 21242 1244 3771 mC.A.mC. A.mC. A. 3772 P.mU. A.fU. A.fU. 0.635 G.mC. A.mU. A.mU. G.fC.fU.G.fU. A.Chl G.fU.mG*mU*mA*mC* mU*mC* U. 21243 1244 3773 mC. A.mC. A.mC.A. 3774 P.mU. A.fU. A.fU. 0.32 G.mC. A.mU. A.mU. G.fC.fU. G.fU. A.ChlG.fU.mG*fU*mA*fC*mU* fC* U. 21244 1244 3775 mC. A.mC. A.mC. A. 3776P.mU. A.fU. A.fU. 0.36 G.mC. A.mU. A.mU. G.fC.fU. G.fU. G.fU. A.ChlG*fU* A*fC*mU*mC* U. 21245 1244 3777 mC. A.mC. A.mC. A. 3778 P.mU. A.fU.A.fU. 0.265 G.mC. A.mU. G.fC.fU. G.fU. G.fU. A*mU*mA.TEG-Chl G*fU*A*fC*fU*fC*fU. 21246 1244 3779 mC*mA*mC. A.mC. 3780 P.mU. A.fU. A.fU.0.334 A. G.mC. A.mU. G.fC.fU. G.fU. G.fU. A*mU*mA.TEG-Chl G*fU*A*fC*fU*fC*fU. 21247 1244 3781 mC*mA*mC.mA.mC. 3782 P.mU. A.fU. A.fU.0.29 mA.mG.mC.mA.mU. G.fC.fU. G.fU. G.fU. mA*mU*mA.TEG-Chl G*fU*A*fC*fU*fC*fU. 21248 614 3783 mA. G. 3784 P.mU.fU. G. A.fC.fU.fU. n/aA.mU.mU.mC. A. A. G. A. A.fU.fC.fU*fC*fU* G.mU.mC*mA*mA.TEG- G*fC*fU* U.Chl 20608 1244 3785 mC. A.mC. A.mC. A. 3786 P.mU. A.fU. A.fU. 79% G.mC.A.mU. A.mU. G.fC.fU. G.fU. G.mU. A.Chl G*mU* A*mC*mU*mC* U. 20609 12443787 mC. A.mC. A.mC. A. 3788 P.mU. A.fU. A.fU. 60% G.mC. A.mU. A.mU.G.fC.fU. G.fU. G.mU. A.Chl G*fU* A*mC*fU*mC* U. 20610 1244 3789 mC.A.mC. A.mC. A. 3790 P.mU. A. U. A. U. G. C. 93% G.mC. A.mU. A.mU. U. G.U. G.mU. G*mU* A.Chl A*mC*mU*mC* U. 20611 1244 3791 mC. A.mC. A.mC. A.3792 P.mU. A.fU. A.fU. n/a G.mC. A.mU. A.mU. G.fC.fU. G.fU. A.ChlG.mU.mG*mU*mA*mC* mU*mC* U. 21374 614 3793 mC*mA*mC.mA.mC. 3794 P.mU.A.fU. A.fU. 24% mA.mG.mC.mA.mU. G.fC.fU. G.fU. mA*mU*mA.TEG-ChlG.fU.mG*fU*mA*fC*mU* fC* U.

TABLE 23 CB1 sequences Ref SEQ ID Pos SEQ ID NO 19-mer Sense Seq NO25-mer Sense Seq w/A @ 25 1690 3795 AUGUCUGUGUCCACAGACA 3796GUAACCAUGUCUGUGUCCACAGACA 1686 3797 AACCAUGUCUGUGUCCACA 3798CAAGGUAACCAUGUCUGUGUCCACA 1685 3799 UAACCAUGUCUGUGUCCAC 3800CCAAGGUAACCAUGUCUGUGUCCAA 1684 3801 GUAACCAUGUCUGUGUCCA 3802GCCAAGGUAACCAUGUCUGUGUCCA 1649 3803 AAAGCUGCAUCAAGAGCAC 3804CCGCAGAAAGCUGCAUCAAGAGCAA 1648 3805 GAAAGCUGCAUCAAGAGCA 3806GCCGCAGAAAGCUGCAUCAAGAGCA 1494 3807 CAUCUAUGCUCUGAGGAGU 3808CCCCAUCAUCUAUGCUCUGAGGAGA 1493 3809 UCAUCUAUGCUCUGAGGAG 3810ACCCCAUCAUCUAUGCUCUGAGGAA 1492 3811 AUCAUCUAUGCUCUGAGGA 3812AACCCCAUCAUCUAUGCUCUGAGGA 1491 3813 CAUCAUCUAUGCUCUGAGG 3814GAACCCCAUCAUCUAUGCUCUGAGA 1490 3815 CCAUCAUCUAUGCUCUGAG 3816UGAACCCCAUCAUCUAUGCUCUGAA 1489 3817 CCCAUCAUCUAUGCUCUGA 3818GUGAACCCCAUCAUCUAUGCUCUGA 1487 3819 ACCCCAUCAUCUAUGCUCU 3820CCGUGAACCCCAUCAUCUAUGCUCA 1486 3821 AACCCCAUCAUCUAUGCUC 3822ACCGUGAACCCCAUCAUCUAUGCUA 1358 3823 UGGUGUUGAUCAUCUGCUG 3824UCCUGGUGGUGUUGAUCAUCUGCUA 1357 3825 GUGGUGUUGAUCAUCUGCU 3826AUCCUGGUGGUGUUGAUCAUCUGCA 1355 3827 UGGUGGUGUUGAUCAUCUG 3828UGAUCCUGGUGGUGUUGAUCAUCUA 1354 3829 CUGGUGGUGUUGAUCAUCU 3830CUGAUCCUGGUGGUGUUGAUCAUCA 1351 3831 AUCCUGGUGGUGUUGAUCA 3832GUCCUGAUCCUGGUGGUGUUGAUCA 1198 3833 AUUCUCUGGAAGGCUCACA 3834AUGUAUAUUCUCUGGAAGGCUCACA 1197 3835 UAUUCUCUGGAAGGCUCAC 3836CAUGUAUAUUCUCUGGAAGGCUCAA 1196 3837 AUAUUCUCUGGAAGGCUCA 3838ACAUGUAUAUUCUCUGGAAGGCUCA 1195 3839 UAUAUUCUCUGGAAGGCUC 3840UACAUGUAUAUUCUCUGGAAGGCUA 1131 3841 CUACCUGAUGUUCUGGAUC 3842UGAAACCUACCUGAUGUUCUGGAUA 1129 3843 ACCUACCUGAUGUUCUGGA 3844GAUGAAACCUACCUGAUGUUCUGGA 1127 3845 AAACCUACCUGAUGUUCUG 3846UUGAUGAAACCUACCUGAUGUUCUA 1126 3847 GAAACCUACCUGAUGUUCU 3848AUUGAUGAAACCUACCUGAUGUUCA 1086 3849 ACUGCAAUCUGUUUGCUCA 3850CGAGAAACUGCAAUCUGUUUGCUCA 1084 3851 AAACUGCAAUCUGUUUGCU 3852UGCGAGAAACUGCAAUCUGUUUGCA 972 3853 CCUGGCCUAUAAGAGGAUU 3854CAGGCCCCUGGCCUAUAAGAGGAUA 951 3855 GUACAUAUCCAUUCACAGG 3856CGACAGGUACAUAUCCAUUCACAGA 950 3857 GGUACAUAUCCAUUCACAG 3858UCGACAGGUACAUAUCCAUUCACAA 948 3859 CAGGUACAUAUCCAUUCAC 3860CAUCGACAGGUACAUAUCCAUUCAA 947 3861 ACAGGUACAUAUCCAUUCA 3862CCAUCGACAGGUACAUAUCCAUUCA 946 3863 GACAGGUACAUAUCCAUUC 3864GCCAUCGACAGGUACAUAUCCAUUA 943 3865 AUCGACAGGUACAUAUCCA 3866ACAGCCAUCGACAGGUACAUAUCCA 941 3867 CCAUCGACAGGUACAUAUC 3868UCACAGCCAUCGACAGGUACAUAUA 940 3869 GCCAUCGACAGGUACAUAU 3870CUCACAGCCAUCGACAGGUACAUAA 869 3871 ACGUGUUUCUGUUCAAACU 3872GCCGCAACGUGUUUCUGUUCAAACA 868 3873 AACGUGUUUCUGUUCAAAC 3874AGCCGCAACGUGUUUCUGUUCAAAA 1647 3875 AGAAAGCUGCAUCAAGAGC 3876GGCCGCAGAAAGCUGCAUCAAGAGA 1645 3877 GCAGAAAGCUGCAUCAAGA 3878AGGGCCGCAGAAAGCUGCAUCAAGA 1394 3879 UCAUGGUGUAUGAUGUCUU 3880UUGCAAUCAUGGUGUAUGAUGUCUA 1393 3881 AUCAUGGUGUAUGAUGUCU 3882CUUGCAAUCAUGGUGUAUGAUGUCA 1391 3883 CAAUCAUGGUGUAUGAUGU 3884UGCUUGCAAUCAUGGUGUAUGAUGA 1125 3885 UGAAACCUACCUGAUGUUC 3886CAUUGAUGAAACCUACCUGAUGUUA 1090 3887 CAAUCUGUUUGCUCAGACA 3888AAACUGCAAUCUGUUUGCUCAGACA 1089 3889 GCAAUCUGUUUGCUCAGAC 3890GAAACUGCAAUCUGUUUGCUCAGAA 1088 3891 UGCAAUCUGUUUGCUCAGA 3892AGAAACUGCAAUCUGUUUGCUCAGA 1087 3893 CUGCAAUCUGUUUGCUCAG 3894GAGAAACUGCAAUCUGUUUGCUCAA 1397 3895 UGGUGUAUGAUGUCUUUGG 3896CAAUCAUGGUGUAUGAUGUCUUUGA 1396 3897 AUGGUGUAUGAUGUCUUUG 3898GCAAUCAUGGUGUAUGAUGUCUUUA 1120 3899 AUUGAUGAAACCUACCUGA 3900CCACACAUUGAUGAAACCUACCUGA 1118 3901 ACAUUGAUGAAACCUACCU 3902UCCCACACAUUGAUGAAACCUACCA 1117 3903 CACAUUGAUGAAACCUACC 3904UUCCCACACAUUGAUGAAACCUACA 1116 3905 ACACAUUGAUGAAACCUAC 3906UUUCCCACACAUUGAUGAAACCUAA 1132 3907 UACCUGAUGUUCUGGAUCG 3908GAAACCUACCUGAUGUUCUGGAUCA 845 3909 UGUUCCACCGCAAAGAUAG 3910UCCACGUGUUCCACCGCAAAGAUAA 844 3911 GUGUUCCACCGCAAAGAUA 3912UUCCACGUGUUCCACCGCAAAGAUA 573 3913 CUUCAAGGAGAAUGAGGAG 3914CUCGUCCUUCAAGGAGAAUGAGGAA 572 3915 CCUUCAAGGAGAAUGAGGA 3916UCUCGUCCUUCAAGGAGAAUGAGGA 571 3917 UCCUUCAAGGAGAAUGAGG 3918CUCUCGUCCUUCAAGGAGAAUGAGA 1449 3919 AUUCUGCAGUAUGCUCUGC 3920GUUUGCAUUCUGCAGUAUGCUCUGA 1448 3921 CAUUCUGCAGUAUGCUCUG 3922UGUUUGCAUUCUGCAGUAUGCUCUA 1447 3923 GCAUUCUGCAGUAUGCUCU 3924GUGUUUGCAUUCUGCAGUAUGCUCA 1253 3925 AGAGCAUCAUCAUCCACAC 3926CCCAGAAGAGCAUCAUCAUCCACAA 1252 3927 AAGAGCAUCAUCAUCCACA 3928ACCCAGAAGAGCAUCAUCAUCCACA 1247 3929 CCCAGAAGAGCAUCAUCAU 3930GUGGCACCCAGAAGAGCAUCAUCAA 1246 3931 ACCCAGAAGAGCAUCAUCA 3932CGUGGCACCCAGAAGAGCAUCAUCA 311 3933 UGAAGUCGAUCCUAGAUGG 3934AGGUUAUGAAGUCGAUCCUAGAUGA 310 3935 AUGAAGUCGAUCCUAGAUG 3936GAGGUUAUGAAGUCGAUCCUAGAUA 1249 3937 CAGAAGAGCAUCAUCAUCC 3938GGCACCCAGAAGAGCAUCAUCAUCA 585 3939 UGAGGAGAACAUCCAGUGU 3940GGAGAAUGAGGAGAACAUCCAGUGA 583 3941 AAUGAGGAGAACAUCCAGU 3942AAGGAGAAUGAGGAGAACAUCCAGA 581 3943 AGAAUGAGGAGAACAUCCA 3944UCAAGGAGAAUGAGGAGAACAUCCA 580 3945 GAGAAUGAGGAGAACAUCC 3946UUCAAGGAGAAUGAGGAGAACAUCA 579 3947 GGAGAAUGAGGAGAACAUC 3948CUUCAAGGAGAAUGAGGAGAACAUA 578 3949 AGGAGAAUGAGGAGAACAU 3950CCUUCAAGGAGAAUGAGGAGAACAA 577 3951 AAGGAGAAUGAGGAGAACA 3952UCCUUCAAGGAGAAUGAGGAGAACA 574 3953 UUCAAGGAGAAUGAGGAGA 3954UCGUCCUUCAAGGAGAAUGAGGAGA 1257 3955 CAUCAUCAUCCACACGUCU 3956GAAGAGCAUCAUCAUCCACACGUCA 1255 3957 AGCAUCAUCAUCCACACGU 3958CAGAAGAGCAUCAUCAUCCACACGA 1682 3959 AGGUAACCAUGUCUGUGUC 3960UUGCCAAGGUAACCAUGUCUGUGUA 1681 3961 AAGGUAACCAUGUCUGUGU 3962AUUGCCAAGGUAACCAUGUCUGUGA 1680 3963 CAAGGUAACCAUGUCUGUG 3964GAUUGCCAAGGUAACCAUGUCUGUA 1499 3965 AUGCUCUGAGGAGUAAGGA 3966UCAUCUAUGCUCUGAGGAGUAAGGA 1498 3967 UAUGCUCUGAGGAGUAAGG 3968AUCAUCUAUGCUCUGAGGAGUAAGA 1497 3969 CUAUGCUCUGAGGAGUAAG 3970CAUCAUCUAUGCUCUGAGGAGUAAA 1496 3971 UCUAUGCUCUGAGGAGUAA 3972CCAUCAUCUAUGCUCUGAGGAGUAA 1388 3973 UUGCAAUCAUGGUGUAUGA 3974CUCUGCUUGCAAUCAUGGUGUAUGA 1387 3975 CUUGCAAUCAUGGUGUAUG 3976CCUCUGCUUGCAAUCAUGGUGUAUA 1386 3977 GCUUGCAAUCAUGGUGUAU 3978CCCUCUGCUUGCAAUCAUGGUGUAA 1385 3979 UGCUUGCAAUCAUGGUGUA 3980GCCCUCUGCUUGCAAUCAUGGUGUA 1384 3981 CUGCUUGCAAUCAUGGUGU 3982GGCCCUCUGCUUGCAAUCAUGGUGA 1383 3983 UCUGCUUGCAAUCAUGGUG 3984GGGCCCUCUGCUUGCAAUCAUGGUA 1382 3985 CUCUGCUUGCAAUCAUGGU 3986GGGGCCCUCUGCUUGCAAUCAUGGA 1314 3987 CCGCAUGGACAUUAGGUUA 3988CCAAGCCCGCAUGGACAUUAGGUUA 1094 3989 CUGUUUGCUCAGACAUUUU 3990UGCAAUCUGUUUGCUCAGACAUUUA 1093 3991 UCUGUUUGCUCAGACAUUU 3992CUGCAAUCUGUUUGCUCAGACAUUA 1083 3993 GAAACUGCAAUCUGUUUGC 3994CUGCGAGAAACUGCAAUCUGUUUGA 1082 3995 AGAAACUGCAAUCUGUUUG 3996ACUGCGAGAAACUGCAAUCUGUUUA 1080 3997 CGAGAAACUGCAAUCUGUU 3998GAACUGCGAGAAACUGCAAUCUGUA 323 3999 UAGAUGGCCUUGCAGAUAC 4000CGAUCCUAGAUGGCCUUGCAGAUAA 322 4001 CUAGAUGGCCUUGCAGAUA 4002UCGAUCCUAGAUGGCCUUGCAGAUA 1179 4003 CGUGUAUGCGUACAUGUAU 4004GUUCAUCGUGUAUGCGUACAUGUAA 1178 4005 UCGUGUAUGCGUACAUGUA 4006UGUUCAUCGUGUAUGCGUACAUGUA 1177 4007 AUCGUGUAUGCGUACAUGU 4008CUGUUCAUCGUGUAUGCGUACAUGA 1320 4009 GGACAUUAGGUUAGCCAAG 4010CCGCAUGGACAUUAGGUUAGCCAAA 1319 4011 UGGACAUUAGGUUAGCCAA 4012CCCGCAUGGACAUUAGGUUAGCCAA 1318 4013 AUGGACAUUAGGUUAGCCA 4014GCCCGCAUGGACAUUAGGUUAGCCA 1317 4015 CAUGGACAUUAGGUUAGCC 4016AGCCCGCAUGGACAUUAGGUUAGCA 1316 4017 GCAUGGACAUUAGGUUAGC 4018AAGCCCGCAUGGACAUUAGGUUAGA 1315 4019 CGCAUGGACAUUAGGUUAG 4020CAAGCCCGCAUGGACAUUAGGUUAA 1415 4021 GGAAGAUGAACAAGCUCAU 4022UCUUUGGGAAGAUGAACAAGCUCAA 552 4023 UUACAACAAGUCUCUCUCG 4024AGAAUUUUACAACAAGUCUCUCUCA 551 4025 UUUACAACAAGUCUCUCUC 4026CAGAAUUUUACAACAAGUCUCUCUA 550 4027 UUUUACAACAAGUCUCUCU 4028ACAGAAUUUUACAACAAGUCUCUCA 476 4029 GUCCCUUCCAAGAGAAGAU 4030GGGGAAGUCCCUUCCAAGAGAAGAA 474 4031 AAGUCCCUUCCAAGAGAAG 4032UAGGGGAAGUCCCUUCCAAGAGAAA 473 4033 GAAGUCCCUUCCAAGAGAA 4034UUAGGGGAAGUCCCUUCCAAGAGAA 1020 4035 UUGCCUGAUGUGGACCAUA 4036GGCGUUUUGCCUGAUGUGGACCAUA 1019 4037 UUUGCCUGAUGUGGACCAU 4038UGGCGUUUUGCCUGAUGUGGACCAA 1018 4039 UUUUGCCUGAUGUGGACCA 4040GUGGCGUUUUGCCUGAUGUGGACCA 606 4041 GGAGAACUUCAUGGACAUA 4042GUGUGGGGAGAACUUCAUGGACAUA 1568 4043 AGCCUCUGGAUAACAGCAU 4044CUGCGCAGCCUCUGGAUAACAGCAA 1170 4045 UCUGUUCAUCGUGUAUGCG 4046ACUGCUUCUGUUCAUCGUGUAUGCA 1169 4047 UUCUGUUCAUCGUGUAUGC 4048UACUGCUUCUGUUCAUCGUGUAUGA 1168 4049 CUUCUGUUCAUCGUGUAUG 4050GUACUGCUUCUGUUCAUCGUGUAUA 1421 4051 UGAACAAGCUCAUUAAGAC 4052GGAAGAUGAACAAGCUCAUUAAGAA 1420 4053 AUGAACAAGCUCAUUAAGA 4054GGGAAGAUGAACAAGCUCAUUAAGA 1419 4055 GAUGAACAAGCUCAUUAAG 4056UGGGAAGAUGAACAAGCUCAUUAAA 1418 4057 AGAUGAACAAGCUCAUUAA 4058UUGGGAAGAUGAACAAGCUCAUUAA 1417 4059 AAGAUGAACAAGCUCAUUA 4060UUUGGGAAGAUGAACAAGCUCAUUA 1172 4061 UGUUCAUCGUGUAUGCGUA 4062UGCUUCUGUUCAUCGUGUAUGCGUA 1078 4063 UGCGAGAAACUGCAAUCUG 4064UGGAACUGCGAGAAACUGCAAUCUA 825 4065 CAGCUUCAUUGACUUCCAC 4066UGUCUACAGCUUCAUUGACUUCCAA 824 4067 ACAGCUUCAUUGACUUCCA 4068UUGUCUACAGCUUCAUUGACUUCCA 823 4069 UACAGCUUCAUUGACUUCC 4070UUUGUCUACAGCUUCAUUGACUUCA 821 4071 UCUACAGCUUCAUUGACUU 4072UUUUUGUCUACAGCUUCAUUGACUA 820 4073 GUCUACAGCUUCAUUGACU 4074AUUUUUGUCUACAGCUUCAUUGACA 612 4075 CUUCAUGGACAUAGAGUGU 4076GGAGAACUUCAUGGACAUAGAGUGA 611 4077 ACUUCAUGGACAUAGAGUG 4078GGGAGAACUUCAUGGACAUAGAGUA 610 4079 AACUUCAUGGACAUAGAGU 4080GGGGAGAACUUCAUGGACAUAGAGA 549 4081 AUUUUACAACAAGUCUCUC 4082UACAGAAUUUUACAACAAGUCUCUA 547 4083 GAAUUUUACAACAAGUCUC 4084AUUACAGAAUUUUACAACAAGUCUA 1176 4085 CAUCGUGUAUGCGUACAUG 4086UCUGUUCAUCGUGUAUGCGUACAUA 1175 4087 UCAUCGUGUAUGCGUACAU 4088UUCUGUUCAUCGUGUAUGCGUACAA 1174 4089 UUCAUCGUGUAUGCGUACA 4090CUUCUGUUCAUCGUGUAUGCGUACA 1173 4091 GUUCAUCGUGUAUGCGUAC 4092GCUUCUGUUCAUCGUGUAUGCGUAA 1171 4093 CUGUUCAUCGUGUAUGCGU 4094CUGCUUCUGUUCAUCGUGUAUGCGA 609 4095 GAACUUCAUGGACAUAGAG 4096UGGGGAGAACUUCAUGGACAUAGAA 608 4097 AGAACUUCAUGGACAUAGA 4098GUGGGGAGAACUUCAUGGACAUAGA 607 4099 GAGAACUUCAUGGACAUAG 4100UGUGGGGAGAACUUCAUGGACAUAA 1322 4101 ACAUUAGGUUAGCCAAGAC 4102GCAUGGACAUUAGGUUAGCCAAGAA 1321 4103 GACAUUAGGUUAGCCAAGA 4104CGCAUGGACAUUAGGUUAGCCAAGA 1027 4105 AUGUGGACCAUAGCCAUUG 4106UGCCUGAUGUGGACCAUAGCCAUUA 545 4107 CAGAAUUUUACAACAAGUC 4108ACAUUACAGAAUUUUACAACAAGUA 532 4109 CAGGUGAACAUUACAGAAU 4110GCAGACCAGGUGAACAUUACAGAAA 813 4111 CAUUUUUGUCUACAGCUUC 4112GAGUGUCAUUUUUGUCUACAGCUUA 812 4113 UCAUUUUUGUCUACAGCUU 4114GGAGUGUCAUUUUUGUCUACAGCUA 811 4115 GUCAUUUUUGUCUACAGCU 4116GGGAGUGUCAUUUUUGUCUACAGCA 809 4117 GUGUCAUUUUUGUCUACAG 4118UGGGGAGUGUCAUUUUUGUCUACAA 808 4119 AGUGUCAUUUUUGUCUACA 4120CUGGGGAGUGUCAUUUUUGUCUACA 569 4121 CGUCCUUCAAGGAGAAUGA 4122CUCUCUCGUCCUUCAAGGAGAAUGA 568 4123 UCGUCCUUCAAGGAGAAUG 4124UCUCUCUCGUCCUUCAAGGAGAAUA 1444 4125 UUUGCAUUCUGCAGUAUGC 4126ACGGUGUUUGCAUUCUGCAGUAUGA 1443 4127 GUUUGCAUUCUGCAGUAUG 4128GACGGUGUUUGCAUUCUGCAGUAUA 1446 4129 UGCAUUCUGCAGUAUGCUC 4130GGUGUUUGCAUUCUGCAGUAUGCUA 1445 4131 UUGCAUUCUGCAGUAUGCU 4132CGGUGUUUGCAUUCUGCAGUAUGCA 1442 4133 UGUUUGCAUUCUGCAGUAU 4134AGACGGUGUUUGCAUUCUGCAGUAA 1677 4135 UGCCAAGGUAACCAUGUCU 4136CAAGAUUGCCAAGGUAACCAUGUCA 1676 4137 UUGCCAAGGUAACCAUGUC 4138UCAAGAUUGCCAAGGUAACCAUGUA 1675 4139 AUUGCCAAGGUAACCAUGU 4140GUCAAGAUUGCCAAGGUAACCAUGA 1603 4141 CUGCACAAACACGCAAACA 4142GACUGCCUGCACAAACACGCAAACA 1110 4143 UUUCCCACACAUUGAUGAA 4144AGACAUUUUCCCACACAUUGAUGAA 1109 4145 UUUUCCCACACAUUGAUGA 4146CAGACAUUUUCCCACACAUUGAUGA 1108 4147 AUUUUCCCACACAUUGAUG 4148UCAGACAUUUUCCCACACAUUGAUA 1605 4149 GCACAAACACGCAAACAAU 4150CUGCCUGCACAAACACGCAAACAAA 1604 4151 UGCACAAACACG CAAACAA 4152ACUGCCUGCACAAACACGCAAACAA 1671 4153 CAAGAUUGCCAAGGUAACC 4154CACGGUCAAGAUUGCCAAGGUAACA 1670 4155 UCAAGAUUGCCAAGGUAAC 4156GCACGGUCAAGAUUGCCAAGGUAAA 1669 4157 GUCAAGAUUGCCAAGGUAA 4158AGCACGGUCAAGAUUGCCAAGGUAA 628 4159 UGUUUCAUGGUCCUGAACC 4160AUAGAGUGUUUCAUGGUCCUGAACA 1115 4161 CACACAUUGAUGAAACCUA 4162UUUUCCCACACAUUGAUGAAACCUA 1114 4163 CCACACAUUGAUGAAACCU 4164AUUUUCCCACACAUUGAUGAAACCA 1113 4165 CCCACACAUUGAUGAAACC 4166CAUUUUCCCACACAUUGAUGAAACA 1112 4167 UCCCACACAUUGAUGAAAC 4168ACAUUUUCCCACACAUUGAUGAAAA 1111 4169 UUCCCACACAUUGAUGAAA 4170GACAUUUUCCCACACAUUGAUGAAA 814 4171 AUUUUUGUCUACAGCUUCA 4172AGUGUCAUUUUUGUCUACAGCUUCA 1659 4173 CAAGAGCACGGUCAAGAUU 4174CUGCAUCAAGAGCACGGUCAAGAUA 1657 4175 AUCAAGAGCACGGUCAAGA 4176AGCUGCAUCAAGAGCACGGUCAAGA 1167 4177 GCUUCUGUUCAUCGUGUAU 4178CGUACUGCUUCUGUUCAUCGUGUAA 1166 4179 UGCUUCUGUUCAUCGUGUA 4180GCGUACUGCUUCUGUUCAUCGUGUA 1668 4181 GGUCAAGAUUGCCAAGGUA 4182GAGCACGGUCAAGAUUGCCAAGGUA 819 4183 UGUCUACAGCUUCAUUGAC 4184CAUUUUUGUCUACAGCUUCAUUGAA 818 4185 UUGUCUACAGCUUCAUUGA 4186UCAUUUUUGUCUACAGCUUCAUUGA 817 4187 UUUGUCUACAGCUUCAUUG 4188GUCAUUUUUGUCUACAGCUUCAUUA 816 4189 UUUUGUCUACAGCUUCAUU 4190UGUCAUUUUUGUCUACAGCUUCAUA 1543 4191 UUUCCCUCUUGUGAAGGCA 4192AGCAUGUUUCCCUCUUGUGAAGGCA 166 4193 AAGAGCACGGUCAAGAUUG 4194UGCAUCAAGAGCACGGUCAAGAUUA 1030 4195 UGGACCAUAGCCAUUGUGA 4196CUGAUGUGGACCAUAGCCAUUGUGA 531 4197 CCAGGUGAACAUUACAGAA 4198AGCAGACCAGGUGAACAUUACAGAA 1259 4199 UCAUCAUCCACACGUCUGA 4200AGAGCAUCAUCAUCCACACGUCUGA 1258 4201 AUCAUCAUCCACACGUCUG 4202AAGAGCAUCAUCAUCCACACGUCUA

Chemical Modification Key Chl cholesterol with hydroxyprolinol linkerTEG-Chl cholesterol with TEG linker m 2′Ome f 2′fluoro *phosphorothioate linkage . phosphodiester linkage

TABLE 24 Summary of CTGF Leads Opt. Tar- lead Opt. Opt. Generic getingse- lead lead Pri- Name TEG ID 2′F site quence Optimized lead sequence2′F 2′OH ority CTGF L1 21045 4 2295 21212 mU.mU. G.mC.A.mC.mC.mU.mU.mU.mC.mU*mA*mA-chol 4  7(2) 1 P.mU.fU. A. G. A. mA. A. G.G.fU. G.fC. mA. mA* mA*fC* mA*mA* mG* G 21214 mU.mU. G.mC.A.mC.mC.mU.mU.mU.mC.mU*mA*mA-chol 4 12(2) 3 P.mU.fU. A. G. A. mA. A.G.fU. G.fC. A. A* A*fC* A*mA*mG*G 21215 mU.mU. G.mC. A.mC.mC.mU.mU.mU.mC.mU*mA*mA-chol 4 10(2) 2 P.mU.fU. A. G. A. mA. A. G.G.fU. G.fC. mA. A* mA*fC* A*mA*mG* G CTGFL2 20393 5 2296 21204 G.mC.A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA-TEG-Chl 3 11(3) 1P.mU.fC.fU.A.G.mA.A.mA.G.G.fU.G.mC*A*A*A*mC*A*U 21205 G.mC.A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA-TEG-Chl 3 10(3) 3P.mU.fC.fC.A.G.mA.A.mA.G.G.fU.G.mC*A*mA*A*mC*A*U 21227 G.mC.A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA-TEG-Chl 5  7(3) 2P.mU.fC.fU.A.G.mA.A.mA.G.G.fU.G.fC*mA*mA*mA*fC*mA*U CTGF L3 20392 132275 21381 G.mU. G. A. mC.mC. A. A. A. A. G*mU*mA-TEG-Chl 6  6(9) 1P.mU.A.fC.fU.fU.fU.fU.G.G.fU.mC.A.mC*A*mC*mU*mC*mU*C 21383mG*mU*mG.mA.mC.mC.mA.mA.mA.mA.mG*mU*mA-TEG-Chl 6  6(0) 2P.mU.A.fC.fU.fU.fU.fU.G.G.fU.mC.A.mC*A*mC*mU*mC*mU*C CTGF L4 17387 52299 21224 mC.mC.mU.mU.mU.mC.mU. A. G.mU.mU* mG*mA-TEG-Chl 5  9(2) 1P.mU.fC. A. A.fC.fU. A. G. A. mA. A. G. G*fU*mG*fC*mA*mA* A Table 24:Lead 21212 corresponds to SEQ ID NOs 3445 and 3446; lead 21214corresponds to SEQ ID NOs 3449 and 3450 (an unmodified form of SEQ IDNO: 3450 corresponds to SEQ ID NO: 4205: UCAACUAGAAAGGUGCAAA); lead21215 corresponds to SEQ ID NOs 3451 and 3452 (an unmodified form of SEQID NO: 3452 corresponds to SEQ ID NO: 4204: UUAGAAAGGUGCAAACAAGG); lead21204 corresponds to SEQ ID NOs 3429 and 3430; lead 21205 corresponds toSEQ ID NOs 3431 and 3432; lead 21227 corresponds to SEQ ID NOs 3475 and3476; lead 21381 corresponds to SEQ ID NOs 3493 and 3494; lead 21383corresponds to SEQ ID NOs 3497 and 3498; and lead 21224 corresponds toSEQ ID NOs 3469 and 3470.

TABLE 25 Summary of PTGS2 Leads Opti- mized Tar- lead Opt. Opt. Genericgeting se- lead lead Pri- Name TEG ID 2′F site quence Optimized leadsequence 2′F 2′OH ority PTGS2 L1 20394 8 448 21228 G. A.mU.mC. A.mC.A.mU.mU.mU. G*mA*mA-TEG-Chl 8  6(5) 1P.mU.fU.fC.A.mA.A.fU.G.fU.G.A.fU.fC*fU*mG*mG*mA*fU* G 21229 G. A.mU.mC.A.mC. A.mU.mU.mU. G*mA*mA-TEG-Chl 4 10(5) 3P.mU.fU.fC.A.A.A.fU.G.fU.G.A.mU.mC*mU*G*G*A*mU*G 21230 G. A.mU.mC. A.mC.A.mU.mU.mU. G*mA*mA-TEG-Chl 8  7(5) 2P.mU.fC.fC.A.A.A.fU.G.fU.G.A.fU.fC*fU*mG*mG*mA*fU* G PTGS2 L2 20395 8449 21293 G. A.mU.mC. A.mC. A.mU.mU.mU. G. A*mU*mA-TEG-Chl 5  8(6) 4P.mU. A.fU.fC. A. A. A.fU. G.fU. G. A.mU.mC*mU*mG*mG*mA*fU* G 21394 G.A.mU.mC. A.mC. A.mU.mU.mU. G. A*mU*mA-TEG-Chl 6 11(6) 3 P.mU. A.fU.fC.A. A. A.fU. G.fU. G. A.mU.fC*mU* G* G* A *fU* G 21233 G. A.mU.mC. A.mC.A. mU.mU.mU. G. A*mU*mA-TEG-Chl 8 11(6) 1P.mU.A.fU.fC.A.A.A.fU.G.fU.G.A.fU.fC*fU*G*G*A*fU*G 21234 G. A.mU.mC.A.mC. A.mU.mU.mU. G. A*mU*mA-TEG-Chl 7  8(6) 2P.mU.A.fU.fC.A.A.A.fU.G.fU.G.A.fU.fC*fU*mG*mG*mA*fU*G Table 25: Lead21228 corresponds to SEQ ID NOs 4309 and 4310; lead 21229 corresponds toSEQ ID NOs 4311 and 4312; lead 21230 corresponds to SEQ ID NOs 4313 and4314; lead 21293 corresponds to SEQ ID NOs 4315 and 4316; lead 21394corresponds to SEQ ID NOs 4317 and 4318; lead 21233 corresponds to SEQID NOs 4319 and 501; and lead 21234 corresponds to SEQ ID NOs 502 and1059.

TABLE 26 Summary of hTGFB1 Leads Optimized Opt. lead Generic NameTargeting site lead sequence Optimized lead sequence Opt. lead 2′F 2′OHPriority TGFb1 hL3 1244 21374 mC*mA*mC.mA.mC.mA.mG.mC. 9 6(0) 1mA.mU.mA*mU*mA-TEG-Chl P.mU.A.fU.A.fU.G.fC.fU.G.fU.G.fU.mG*fU*mA*fC*mU*fC*U Table 26: lead 21374 corresponds to SEQ ID NOs3793 and 3794.

TABLE 27 Summary of hTGFB2 Leads Generic Targeting Optimized Opt. leadOpt. lead Name site lead sequence Optimized lead sequence 2′F 2′OHPriority TGFb2 hL3 2081 21379 mU*mC*mA.mU.mC.mA.mG.mU.mG. 7 9(0) 1mU.mU*mA-TEG-Chl P.mU.fU. A. A.fC. A.fC.fU. G. A.fU. G.A* A*fC*fC*mA*mA* G mU*mC*mA.mU.mC.mA.mG.mU.mG. 7 7(0) 2mU.mU*mA*mA-TEG-Chl P.mU.fU. A. A.fC. A.fC.fU. G. A.fU. G.mA*mA*fC*fC*mA*mA* G Table 27: lead 21379 corresponds to SEQ ID NOs3637, 3638, 3639 and 3640.

Having thus described several aspects of at least one embodiment of thisinvention, it is to be appreciated various alterations, modifications,and improvements will readily occur to those skilled in the art. Suchalterations, modifications, and improvements are intended to be part ofthis disclosure, and are intended to be within the spirit and scope ofthe invention. Accordingly, the foregoing description and drawings areby way of example only.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

All references, including patent documents, disclosed herein areincorporated by reference in their entirety. This applicationincorporates by reference the entire contents, including all thedrawings and all parts of the specification (including sequence listingor amino acid/polynucleotide sequences) of PCT Publication No.WO2010/033247 (Application No. PCT/US2009/005247), filed on Sep. 22,2009, and entitled “REDUCED SIZE SELF-DELIVERING RNAI COMPOUNDS” and PCTPublication No. WO2009/102427 (Application No. PCT/US2009/000852), filedon Feb. 11, 2009, and entitled, “MODIFIED RNAI POLYNUCLEOTIDES AND USESTHEREOF.”

What is claimed is:
 1. A method for treating or preventing a fibroticdisorder, the method comprising administering to a subject in needthereof a therapeutically effective amount of a double-strandedribonucleic acid (dsRNA) directed against CTGF comprising a sense strandand an antisense strand, wherein the sense strand comprises at least 12contiguous nucleotides of a sequence selected from the group consistingof SEQ ID NOs: 2463 (GCACCUUUCUAGA), 3429 (G.mC.A.mC.mC.mU.mU.mU.mC.mU.A*mG*mA.TEG-Chl), 2443 (UUGCACCUUUCUAA), 3445 (mU.mU.G.mC.A.mC.mC.mU.mU.mU.mC.mU*mA*mA.TEG-Chl), 2465 (CCUUUCUAGUUGA),

and 3469 (mC.mC.mU.mU.mU.mC.mU.A.G.mU.mU*mG*mA.TEG-Chi)

and/or wherein the antisense strand comprises at least 12 contiguousnucleotides of a sequence selected from the group consisting ofSEQ ID NOs: 2464 (UCUAGAAAGGUGCAAACAU), 3430 (P.mU.fC.fU.A.G.mA.A.mA.G.G.fU.G.mC*A*A*A*mC*A*U), 4203(UUAGAAAGGUGCAAACAAGG), 3446 (P.mU.fU.A.G.A.mA.A.G.G.fU.G.fC.mA.mA*mA*fC*mA*mA*mG* G.), 2466 (UCAACUAGAAAGGUGCAAA),

and 3470 (P.mU.fC.A.A.fC.fU.A.G.A.mA.A.G.G* fU*mG*fC*mA*mA*A.),

wherein the antisense strand is 16-23 nucleotides long and the sensestrand is 8-15 nucleotides long, wherein the sd-rxRNA includes adouble-stranded region and a single-stranded region, wherein thedouble-stranded region is from 8-15 nucleotides long, wherein thesingle-stranded region is at the 3′ end of the antisense strand and is4-12 nucleotides long, wherein the single-stranded region contains 3, 4,5, 6, 7, 8, 9, 10, 11 or 12 phosphorothioate modifications, and whereinat least 40% of the nucleotides of the isolated double-stranded nucleicacid molecule are modified.
 2. The method of claim 1, wherein the dsRNAis administered via intradermal injection.
 3. The method of claim 1,wherein the dsRNA is administered locally to the skin.
 4. The method ofclaim 1, wherein two or more dsRNAs are administered simultaneously orsequentially.
 5. The method of claim 1, wherein the dsRNA ishydrophobically modified.
 6. The method of claim 1, wherein the dsRNA islinked to a hydrophobic conjugate.
 7. The method of claim 1, wherein thesense strand comprises SEQ ID NO: 2463 (GCACCUUUCUAGA)

and the antisense strand comprises SEQ ID NO: 2464(UCUAGAAAGGUGCAAACAU).


8. The method of claim 7, wherein the sense strand comprisesSEQ ID NO: 3429 (G.mC. A.mC.mC.mU.mU.mU.mC.mU. A*mG*mA.TEG-Chl)

and the antisense strand comprises SEQ ID NO: 3430(P.mU.fC.fU. A. G.mA. A.mA. G. G.fU. G.mC* A* A* A*mC* A* U).


9. The method of claim 1, wherein the sense strand comprisesSEQ ID NO: 2443 (UUGCACCUUUCUAA)

and the antisense strand comprises SEQ ID NO: 4203(UUAGAAAGGUGCAAACAAGG).


10. The method of claim 9, wherein the sense strand comprisesSEQ ID NO: 3445 (mU.mU.G.mC.A.mC.mC.mU.mU.mU.mC.mU*mA* mA.TEG-Chl)

and the antisense strand comprises SEQ ID NO: 3446(P.mU.fU. A. G. A.mA. A. G. G.fU. G.fC.mA.mA*mA*fC*mA*mA*mG* G.).


11. The method of claim 1, wherein the sense strand comprisesSEQ ID NO: 2465 (CCUUUCUAGUUGA)

and the antisense strand comprises SEQ ID NO: 2466(UCAACUAGAAAGGUGCAAA).


12. The method of claim 11, wherein the sense strand comprisesSEQ ID NO: 3469 (mC.mC.mU.mU.mU.mC.mU. A. G.mU.mU*mG*mA.TEG-Chl)

and the antisense strand comprises SEQ ID NO: 3470(P.mU.fC. A. A.fC.fU. A. G. A.mA. A. G. G*fU*mG*fC*mA*mA* A.).


13. The method of claim 1, wherein the fibrotic disorder is selectedfrom the group consisting of pulmonary fibrosis, liver cirrhosis,scleroderma and glomerulonephritis, lung fibrosis, liver fibrosis, skinfibrosis, muscle fibrosis, radiation fibrosis, kidney fibrosis,proliferative vitreoretinopathy, restenosis and uterine fibrosis, andtrabeculectomy failure due to scarring.
 14. The method of claim 1wherein the dsRNA is formulated for delivery to the skin, for topicaldelivery, for intradermal injection and/or wherein the dsRNA is in aneutral formulation.
 15. A method for treating or preventing a fibroticdisorder, the method comprising administering to a subject in needthereof a therapeutically effective amount of a double-strandedribonucleic acid (dsRNA) directed against CTGF comprising a sense strandand an antisense strand, wherein the sense strand comprises at least 12contiguous nucleotides of a sequence selected from the group consistingof SEQ ID NOs: 2459 (GUGACCAAAAGUA) or 3493(G.mU. G. A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl)

and/or wherein the antisense strand comprises at least 12 contiguousnucleotides of SEQ ID NOs: 2460 (UACUUUUGGUCACACUCUC) or 3494P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC*A*mC*mU*mC*mU* C.),

wherein the dsRNA is an sd-rxRNA, wherein the antisense strand is 16-23nucleotides long and the sense strand is 8-15 nucleotides long, whereinthe sd-rxRNA includes a double-stranded region and a single-strandedregion, wherein the double-stranded region is from 8-15 nucleotideslong, wherein the single-stranded region is at the 3′ end of theantisense strand and is 4-12 nucleotides long, wherein thesingle-stranded region contains 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12phosphorothioate modifications, and wherein at least 40% of thenucleotides of the isolated double-stranded nucleic acid molecule aremodified.
 16. The method of claim 15, wherein the sense strand comprisesSEQ ID NO: 2459 (GUGACCAAAAGUA)

and the antisense strand comprises SEQ ID NO: 2460(UACUUUUGGUCACACUCUC),

or the sense strand comprises SEQ ID NO: 3493(G.mU. G.A.mC.mC. A. A. A. A. G*mU*mA.TEG-Chl)

and the antisense strand comprises SEQ ID NO: 3494(P.mU. A.fC.fU.fU.fU.fU. G. G.fU.mC. A.mC*  A*mC*mU*mC*mU* C.).


17. The method of claim 14, wherein the dsRNA is administered viaintradermal injection.
 18. The method of claim 14, wherein the dsRNA isadministered locally to the skin.
 19. The method of claim 14, whereindsRNA is hydrophobically modified.
 20. The method of claim 14, whereinthe dsRNA is linked to a hydrophobic conjugate.